Richmond Ltd. has recently undergone a change in strategy, away from the traditional commodity based meat industry, towards the modern food business. To do this, opportunities to add value to their current product range must be identified. This involves the conversion of traditionally low value commodity based products into products that demand a premium. An example of this is converting muscles that are currently used for grinding meat into a further processed convenience food (i.e. ready meals). Another method is to add further value to premium products by making them more appealing to consumers (i.e. nutritional information on labels). This work details investigations into the functional properties of selected beef forequarter muscles (low value commodity products) and the nutritional properties of selected export lamb products (premium products). The functional properties of a number of beef forequarter muscles were measured to identify which had the best potential for further processing applications with respect to ready meals. The functional properties of tenderness, cook loss and shrinkage were measured for the Latissimus Dorsi, Pectorialis Profundus (Point End Brisket), Infraspinatus (Cross Cut Blade), Triceps Brachi Longhead (Main muscle in Bolar Shoulder Clod), Supraspinatus (Chuck Tender), Serratus Ventralis and Triceps Brachi Medialhead (Muscle in Bolar Shoulder Clod. From the tests conducted the Infraspinatus and the Triceps Brachi Longhead have been identified as having the best functional properties with respect to further processing for ready meal applications. As well as conducting tests to identify the forequarter muscles with the best potential for further processing applications, investigations were carried out to identify cooking regimes that would optimise the functional properties. This work confirmed that there are three major chemical reactions, which determine the resultant functional properties of cooked meat. They are the denaturation and aggregation of the myofibrillar proteins and the denaturation and solubilisation of connective tissue (collagen). At around 50°C myosin (45% to 50% of the myofibrillar proteins) denatures, which results in a substantial increase in cook loss and reduction in water holding capacity. At around 60°C collagen (main connective tissue protein) denatures, which results in a substantial increase in tenderness and increase in cook loss. This is because as the collagen denatures it loses it mechanical strength (increase in tenderness) and can no longer support its own structure, and causes it to contract. This contraction causes fluid within the meat and cook loss caused by the denaturation of myosin to be expelled from the meat by compressive forces (squeezed out). At around 70°C actomyosin (22% of the myofibrillar proteins) denatures. This results in a substantial increase in the cook loss and firming of the meat. The increase in cook loss or decrease in water holding capacity that occurs with myofibrillar protein denaturation is due to the fact that when these proteins denature and aggregate their ability to bind water is greatly reduced. From the results of the cooking regime trials it is recommended that for functional property considerations that during the cooking of further processed meat products (i.e. ready meal applications) a meat temperature of 62°C should be aimed for, for the slowest heating region during cooking (usually the centre). This is because it has been identified that a cooking temperature of 65°C should not be exceeded otherwise detrimental effects can occur to the functional properties of the cooked meat. For health concerns a 7D bacterial death reduction has to be achieved. This means that for a cooking temperature of 62°C the meat has to be held at this temperature for at least 5 minutes. Therefore the total cooking time would be the time needed to heat all the meat to 62°C plus 5 minutes to ensure a safe product. The heating or cooking system employed should also ensure that a minimal amount of the meat is heated above 65°C. This can be easily achieved by minimising the external cooking temperature, but long cooking times will result. An industrial cooking process will be a compromise between the cost associated with longer residence time and product functionality. As mentioned earlier another way to add value is to supply nutritional information for selected cuts. Consequentially one of the objectives of this project was to provide some nutritional information for selected meat cuts. Though the primary objective of this part of the project was to develop a method for producing the needed information, so that Richmond N.Z. Ltd. can develop further information on an as needs basis. The nutritional characteristics of a number of export lamb cuts from the saddle region has also been investigated and a method devised to allow further characterisation of other cuts. The method involves breaking down a standard cut into its constituent components (e.g. Frenched rack consists of loin eye, fat cap, intercostals and fatty tissue). The constituent components are tested for their nutritional properties. The frenched rack nutritional properties are calculated from the nutritional properties of the constituents components and the yield data (percentage of each constituent component within a frenched rack) for frenched racks. This method allowed the identification of the main sources of variation for nutritional characteristics. These differences were found to be caused by the lean to fat ratio, not nutritional differences in lean tissue from the same region of lamb (i.e. loin eye and tenderloin very similar nutritionally). The difference in lean to fat ration also accounts for the variation between grades (i.e. PX grade lamb cuts have a higher fat content than YX grade lamb cuts due to PX grade cuts having a higher percentage fat tissue in their cuts). The cuts characterised were the shortloin section (whole section or chop), rack section (whole section or chop), 75mm racks frenched 25mm, boneless loin and tenderloin for both PX and YX grade lamb. The method will be applicable to other regions of lamb (i.e. hindquarter and forequarter) for which nutritional information already exists, but for which yielding data will have to be collected. The method would also be applicable to other species such as beef and venison, but both nutritional data for constituent components and yielding data would have to be collected.

Richmond Ltd. has recently undergone a change in strategy, away from the traditional commodity based meat industry, towards the modern food business. To do this, opportunities to add value to their current product range must be identified. This involves the conversion of traditionally low value commodity based products into products that demand a premium. An example of this is converting muscles that are currently used for grinding meat into a further processed convenience food (i.e. ready meals). Another method is to add further value to premium products by making them more appealing to consumers (i.e. nutritional information on labels). This work details investigations into the functional properties of selected beef forequarter muscles (low value commodity products) and the nutritional properties of selected export lamb products (premium products). The functional properties of a number of beef forequarter muscles were measured to identify which had the best potential for further processing applications with respect to ready meals. The functional properties of tenderness, cook loss and shrinkage were measured for the Latissimus Dorsi, Pectorialis Profundus (Point End Brisket), Infraspinatus (Cross Cut Blade), Triceps Brachi Longhead (Main muscle in Bolar Shoulder Clod), Supraspinatus (Chuck Tender), Serratus Ventralis and Triceps Brachi Medialhead (Muscle in Bolar Shoulder Clod. From the tests conducted the Infraspinatus and the Triceps Brachi Longhead have been identified as having the best functional properties with respect to further processing for ready meal applications. As well as conducting tests to identify the forequarter muscles with the best potential for further processing applications, investigations were carried out to identify cooking regimes that would optimise the functional properties. This work confirmed that there are three major chemical reactions, which determine the resultant functional properties of cooked meat. They are the denaturation and aggregation of the myofibrillar proteins and the denaturation and solubilisation of connective tissue (collagen). At around 50°C myosin (45% to 50% of the myofibrillar proteins) denatures, which results in a substantial increase in cook loss and reduction in water holding capacity. At around 60°C collagen (main connective tissue protein) denatures, which results in a substantial increase in tenderness and increase in cook loss. This is because as the collagen denatures it loses it mechanical strength (increase in tenderness) and can no longer support its own structure, and causes it to contract. This contraction causes fluid within the meat and cook loss caused by the denaturation of myosin to be expelled from the meat by compressive forces (squeezed out). At around 70°C actomyosin (22% of the myofibrillar proteins) denatures. This results in a substantial increase in the cook loss and firming of the meat. The increase in cook loss or decrease in water holding capacity that occurs with myofibrillar protein denaturation is due to the fact that when these proteins denature and aggregate their ability to bind water is greatly reduced. From the results of the cooking regime trials it is recommended that for functional property considerations that during the cooking of further processed meat products (i.e. ready meal applications) a meat temperature of 62°C should be aimed for, for the slowest heating region during cooking (usually the centre). This is because it has been identified that a cooking temperature of 65°C should not be exceeded otherwise detrimental effects can occur to the functional properties of the cooked meat. For health concerns a 7D bacterial death reduction has to be achieved. This means that for a cooking temperature of 62°C the meat has to be held at this temperature for at least 5 minutes. Therefore the total cooking time would be the time needed to heat all the meat to 62°C plus 5 minutes to ensure a safe product. The heating or cooking system employed should also ensure that a minimal amount of the meat is heated above 65°C. This can be easily achieved by minimising the external cooking temperature, but long cooking times will result. An industrial cooking process will be a compromise between the cost associated with longer residence time and product functionality. As mentioned earlier another way to add value is to supply nutritional information for selected cuts. Consequentially one of the objectives of this project was to provide some nutritional information for selected meat cuts. Though the primary objective of this part of the project was to develop a method for producing the needed information, so that Richmond N.Z. Ltd. can develop further information on an as needs basis. The nutritional characteristics of a number of export lamb cuts from the saddle region has also been investigated and a method devised to allow further characterisation of other cuts. The method involves breaking down a standard cut into its constituent components (e.g. Frenched rack consists of loin eye, fat cap, intercostals and fatty tissue). The constituent components are tested for their nutritional properties. The frenched rack nutritional properties are calculated from the nutritional properties of the constituents components and the yield data (percentage of each constituent component within a frenched rack) for frenched racks. This method allowed the identification of the main sources of variation for nutritional characteristics. These differences were found to be caused by the lean to fat ratio, not nutritional differences in lean tissue from the same region of lamb (i.e. loin eye and tenderloin very similar nutritionally). The difference in lean to fat ration also accounts for the variation between grades (i.e. PX grade lamb cuts have a higher fat content than YX grade lamb cuts due to PX grade cuts having a higher percentage fat tissue in their cuts). The cuts characterised were the shortloin section (whole section or chop), rack section (whole section or chop), 75mm racks frenched 25mm, boneless loin and tenderloin for both PX and YX grade lamb. The method will be applicable to other regions of lamb (i.e. hindquarter and forequarter) for which nutritional information already exists, but for which yielding data will have to be collected. The method would also be applicable to other species such as beef and venison, but both nutritional data for constituent components and yielding data would have to be collected.

The objective of this study was to investigate the effects of preheat treatments on fouling by fresh whole milk (FWM), recombined whole milk (RCB) and reconstituted whole milk (Recon) in the high-temperature heater of indirect UHT plants. Various preheat treatments prior to evaporation during milk powder manufacture were applied to skim milk powder (SMP, 75 °C 2 s, 85 °C, 155 s and 95 °C, 155 s) and whole milk powder (WMP, 95 °C, 33 s). These preheat treatments were so-called “evaporator preheat treatments”. Skim milk powder (SMP) and whole milk powder (WMP) were derived from the same original batch of pasteurised FWM to remove the effects of the variation in milk composition between different milk batches. These SMPs were recombined with anhydrous milk fat and water to prepare RCB, and WMPs were reconstituted with water to prepare Recon. Then, (homogenized) FWM, RCB and Recon were subjected to various preheat treatments (75 °C, 11 s, 85 °C, 147 s and 95 °C, 147 s) prior to UHT processing. These preheat treatments were so-called “UHT preheat treatments”. Temperature difference (hot water inlet temperature – milk outlet temperature) was taken as a measure of the extent of fouling in the high-temperature heater. The slope of the linear regression of temperature difference versus time (for two hours of UHT processing) was taken as fouling rate (°C/h). Increasing both evaporator and UHT preheat treatments resulted in increasing fouling rate and total deposit weight for all three whole milk types for several milk batches. In the case of FWM, there was no reduction in fouling rate with increasing UHT preheat treatment whether FWM was homogenized then preheated, preheated then homogenized or not homogenized at all. These findings, which are wholly consistent and well replicated, are in apparent conflict with the results of most previous comparable studies. Possible reasons for this are explained. Further investigations of the effects of homogenization relating to the role of whey protein on the surface of the fat globules showed that whey protein associated with the membrane covering the surface of fat globules for homogenized then preheated FWM, RCB and Recon and that association increased with increasing heating process stage. The increasing association of whey protein with the milk fat globules membrane with increasing severity of heating process stage became faster when preheat treatment was more severe: the association of whey protein plateaued on intermediate temperature heating when the milks were preheated at 75°C, 11 s and on preheating when the milks were preheated at 95°C, 147 s. In the case of FWM, the thickness of the membrane covering the surface of fat globules for homogenized then preheated FWM, which increased with the severity of heating process stage, was greater than the thickness of the membrane in preheated then homogenized FWM. Preheating then homogenization resulted in the greater interfacial spreading of small molecules on the surface of fat globules, i.e. whey protein or small molecules from the disintegration of casein micelles during preheating. Possible basic mechanisms for UHT fouling in the high-temperature heater include: the reduction in the solubility of calcium phosphate and the deposition of protein as fat-bound protein and non-fat-bound protein. When non-fat-bound protein in milk plasma deposited, it could be a carrier for the deposition of mineral, such as, the precipitate of calcium phosphate in the casein micelles or the deposition of complexes between whey protein and casein micelles.

Richmond Ltd. has recently undergone a change in strategy, away from the traditional commodity based meat industry, towards the modern food business. To do this, opportunities to add value to their current product range must be identified. This involves the conversion of traditionally low value commodity based products into products that demand a premium. An example of this is converting muscles that are currently used for grinding meat into a further processed convenience food (i.e. ready meals). Another method is to add further value to premium products by making them more appealing to consumers (i.e. nutritional information on labels). This work details investigations into the functional properties of selected beef forequarter muscles (low value commodity products) and the nutritional properties of selected export lamb products (premium products). The functional properties of a number of beef forequarter muscles were measured to identify which had the best potential for further processing applications with respect to ready meals. The functional properties of tenderness, cook loss and shrinkage were measured for the Latissimus Dorsi, Pectorialis Profundus (Point End Brisket), Infraspinatus (Cross Cut Blade), Triceps Brachi Longhead (Main muscle in Bolar Shoulder Clod), Supraspinatus (Chuck Tender), Serratus Ventralis and Triceps Brachi Medialhead (Muscle in Bolar Shoulder Clod. From the tests conducted the Infraspinatus and the Triceps Brachi Longhead have been identified as having the best functional properties with respect to further processing for ready meal applications. As well as conducting tests to identify the forequarter muscles with the best potential for further processing applications, investigations were carried out to identify cooking regimes that would optimise the functional properties. This work confirmed that there are three major chemical reactions, which determine the resultant functional properties of cooked meat. They are the denaturation and aggregation of the myofibrillar proteins and the denaturation and solubilisation of connective tissue (collagen). At around 50°C myosin (45% to 50% of the myofibrillar proteins) denatures, which results in a substantial increase in cook loss and reduction in water holding capacity. At around 60°C collagen (main connective tissue protein) denatures, which results in a substantial increase in tenderness and increase in cook loss. This is because as the collagen denatures it loses it mechanical strength (increase in tenderness) and can no longer support its own structure, and causes it to contract. This contraction causes fluid within the meat and cook loss caused by the denaturation of myosin to be expelled from the meat by compressive forces (squeezed out). At around 70°C actomyosin (22% of the myofibrillar proteins) denatures. This results in a substantial increase in the cook loss and firming of the meat. The increase in cook loss or decrease in water holding capacity that occurs with myofibrillar protein denaturation is due to the fact that when these proteins denature and aggregate their ability to bind water is greatly reduced. From the results of the cooking regime trials it is recommended that for functional property considerations that during the cooking of further processed meat products (i.e. ready meal applications) a meat temperature of 62°C should be aimed for, for the slowest heating region during cooking (usually the centre). This is because it has been identified that a cooking temperature of 65°C should not be exceeded otherwise detrimental effects can occur to the functional properties of the cooked meat. For health concerns a 7D bacterial death reduction has to be achieved. This means that for a cooking temperature of 62°C the meat has to be held at this temperature for at least 5 minutes. Therefore the total cooking time would be the time needed to heat all the meat to 62°C plus 5 minutes to ensure a safe product. The heating or cooking system employed should also ensure that a minimal amount of the meat is heated above 65°C. This can be easily achieved by minimising the external cooking temperature, but long cooking times will result. An industrial cooking process will be a compromise between the cost associated with longer residence time and product functionality. As mentioned earlier another way to add value is to supply nutritional information for selected cuts. Consequentially one of the objectives of this project was to provide some nutritional information for selected meat cuts. Though the primary objective of this part of the project was to develop a method for producing the needed information, so that Richmond N.Z. Ltd. can develop further information on an as needs basis. The nutritional characteristics of a number of export lamb cuts from the saddle region has also been investigated and a method devised to allow further characterisation of other cuts. The method involves breaking down a standard cut into its constituent components (e.g. Frenched rack consists of loin eye, fat cap, intercostals and fatty tissue). The constituent components are tested for their nutritional properties. The frenched rack nutritional properties are calculated from the nutritional properties of the constituents components and the yield data (percentage of each constituent component within a frenched rack) for frenched racks. This method allowed the identification of the main sources of variation for nutritional characteristics. These differences were found to be caused by the lean to fat ratio, not nutritional differences in lean tissue from the same region of lamb (i.e. loin eye and tenderloin very similar nutritionally). The difference in lean to fat ration also accounts for the variation between grades (i.e. PX grade lamb cuts have a higher fat content than YX grade lamb cuts due to PX grade cuts having a higher percentage fat tissue in their cuts). The cuts characterised were the shortloin section (whole section or chop), rack section (whole section or chop), 75mm racks frenched 25mm, boneless loin and tenderloin for both PX and YX grade lamb. The method will be applicable to other regions of lamb (i.e. hindquarter and forequarter) for which nutritional information already exists, but for which yielding data will have to be collected. The method would also be applicable to other species such as beef and venison, but both nutritional data for constituent components and yielding data would have to be collected.

The increasing price of oil and the destabilisation of the world’s climate are urging governments, businesses and individuals to constantly investigate energy-efficient technologies and methodologies and pursue the adoption of energy efficiency programmes in a global effort to reduce energy consumption, greenhouse gas emissions and ultimately energy costs. In New Zealand, one of the biggest industrial energy efficiency projects was started in 2002 by a multinational dairy company, the Fonterra Co-operative Group, in partnership with the energy service company Demand Response Ltd; the project currently aims at reducing by 15% the energy costs at all Fonterra’s major production sites throughout the country. This thesis, undertaken as part of the above project, examines the development and implementation of a structured and integrated energy monitoring and targeting methodology (M&T) for the most efficient operation of all Fonterra’s chilled water systems, with an initial focus on the ones installed at Clandeboye, one of the Fonterra’s sites involved in the energy saving project. A data collection system (Insite) was already in place at Clandeboye to enable storage and analysis of some of the site’s utility metering data. After identification of key chilled water system components and definition of data requirements for M&T purposes, an analysis of past energy consumption trends (based on multiple regression calculations) was carried out to develop an historical benchmark of the energy used, compare it with current energy performance and thus identify opportunities for future improvements. The creation of an M&T reporting system for presenting findings to operators and management was the last essential part of the thesis development. The study has highlighted that the robustness of the proposed regression model was badly affected by the unreliability of the existing data collection system and the uncertainty associated with poorly documented changes to operating conditions/plant configuration that had occurred over time. The conclusion is that, while the developed M&T methodology is theoretically valid and readily applicable, further developments are necessary (and recommended) to make it suitable for other similar systems.

Richmond Ltd. has recently undergone a change in strategy, away from the traditional commodity based meat industry, towards the modern food business. To do this, opportunities to add value to their current product range must be identified. This involves the conversion of traditionally low value commodity based products into products that demand a premium. An example of this is converting muscles that are currently used for grinding meat into a further processed convenience food (i.e. ready meals). Another method is to add further value to premium products by making them more appealing to consumers (i.e. nutritional information on labels). This work details investigations into the functional properties of selected beef forequarter muscles (low value commodity products) and the nutritional properties of selected export lamb products (premium products). The functional properties of a number of beef forequarter muscles were measured to identify which had the best potential for further processing applications with respect to ready meals. The functional properties of tenderness, cook loss and shrinkage were measured for the Latissimus Dorsi, Pectorialis Profundus (Point End Brisket), Infraspinatus (Cross Cut Blade), Triceps Brachi Longhead (Main muscle in Bolar Shoulder Clod), Supraspinatus (Chuck Tender), Serratus Ventralis and Triceps Brachi Medialhead (Muscle in Bolar Shoulder Clod. From the tests conducted the Infraspinatus and the Triceps Brachi Longhead have been identified as having the best functional properties with respect to further processing for ready meal applications. As well as conducting tests to identify the forequarter muscles with the best potential for further processing applications, investigations were carried out to identify cooking regimes that would optimise the functional properties. This work confirmed that there are three major chemical reactions, which determine the resultant functional properties of cooked meat. They are the denaturation and aggregation of the myofibrillar proteins and the denaturation and solubilisation of connective tissue (collagen). At around 50°C myosin (45% to 50% of the myofibrillar proteins) denatures, which results in a substantial increase in cook loss and reduction in water holding capacity. At around 60°C collagen (main connective tissue protein) denatures, which results in a substantial increase in tenderness and increase in cook loss. This is because as the collagen denatures it loses it mechanical strength (increase in tenderness) and can no longer support its own structure, and causes it to contract. This contraction causes fluid within the meat and cook loss caused by the denaturation of myosin to be expelled from the meat by compressive forces (squeezed out). At around 70°C actomyosin (22% of the myofibrillar proteins) denatures. This results in a substantial increase in the cook loss and firming of the meat. The increase in cook loss or decrease in water holding capacity that occurs with myofibrillar protein denaturation is due to the fact that when these proteins denature and aggregate their ability to bind water is greatly reduced. From the results of the cooking regime trials it is recommended that for functional property considerations that during the cooking of further processed meat products (i.e. ready meal applications) a meat temperature of 62°C should be aimed for, for the slowest heating region during cooking (usually the centre). This is because it has been identified that a cooking temperature of 65°C should not be exceeded otherwise detrimental effects can occur to the functional properties of the cooked meat. For health concerns a 7D bacterial death reduction has to be achieved. This means that for a cooking temperature of 62°C the meat has to be held at this temperature for at least 5 minutes. Therefore the total cooking time would be the time needed to heat all the meat to 62°C plus 5 minutes to ensure a safe product. The heating or cooking system employed should also ensure that a minimal amount of the meat is heated above 65°C. This can be easily achieved by minimising the external cooking temperature, but long cooking times will result. An industrial cooking process will be a compromise between the cost associated with longer residence time and product functionality. As mentioned earlier another way to add value is to supply nutritional information for selected cuts. Consequentially one of the objectives of this project was to provide some nutritional information for selected meat cuts. Though the primary objective of this part of the project was to develop a method for producing the needed information, so that Richmond N.Z. Ltd. can develop further information on an as needs basis. The nutritional characteristics of a number of export lamb cuts from the saddle region has also been investigated and a method devised to allow further characterisation of other cuts. The method involves breaking down a standard cut into its constituent components (e.g. Frenched rack consists of loin eye, fat cap, intercostals and fatty tissue). The constituent components are tested for their nutritional properties. The frenched rack nutritional properties are calculated from the nutritional properties of the constituents components and the yield data (percentage of each constituent component within a frenched rack) for frenched racks. This method allowed the identification of the main sources of variation for nutritional characteristics. These differences were found to be caused by the lean to fat ratio, not nutritional differences in lean tissue from the same region of lamb (i.e. loin eye and tenderloin very similar nutritionally). The difference in lean to fat ration also accounts for the variation between grades (i.e. PX grade lamb cuts have a higher fat content than YX grade lamb cuts due to PX grade cuts having a higher percentage fat tissue in their cuts). The cuts characterised were the shortloin section (whole section or chop), rack section (whole section or chop), 75mm racks frenched 25mm, boneless loin and tenderloin for both PX and YX grade lamb. The method will be applicable to other regions of lamb (i.e. hindquarter and forequarter) for which nutritional information already exists, but for which yielding data will have to be collected. The method would also be applicable to other species such as beef and venison, but both nutritional data for constituent components and yielding data would have to be collected.

Agglomerated milk powders are produced to give improved properties such as flowability, dispersibility, reduced dustiness and decreased bulk density. A key function of these powders is to dissolve "instantly" upon addition to water and because of this they are also called "instant milk powders". They are produced by agglomerating the undersized fines that are returned to the top of the spray drier with milk concentrate droplet spray. Interaction occurs in a collision zone, often with multiple sprays and fines return lines. Agglomeration can be a difficult process to control and operators find it hard to fine tune the process to produce specific powder properties. This work aimed to understand the effects of key droplet and fines properties on the extent of agglomeration to allow a mechanistic understanding of the process. Three scales of spray drier were investigated in this study with different rates of evaporation; a small scale drier (0.5 - 7 kg water h-1), a pilot scale drier (80 kg water h-1) and a range of commercial production scale driers (4 - 15 000 kg water h-1). A survey of operators of commercial scale driers showed that control of instant milk powder production to influence bulk density is highly intuitive. Fines recycle rates were expected to be important in control of agglomeration processes and were estimated on a specific plant by using the pressure drop measured in the fines return line. A model based on pressure drop along a pneumatic pipeline under-predicted the experimental values for pressure drop due to solids, which means a calibration curve should be generated for each specific drier. Fines recycle rates were predicted to be significantly higher at 95 to 130 % of production rates compared to those expected by operators of 50%. Experimental measurements agreed with existing models for the effect of temperature on the density and viscosity of milk concentrates. Experimental results showed that the surface tensions of concentrated milks were within the same range as literature values for standard milks below 60°C, but were significantly higher for milk above 60°C. This is thought to be linked to the mechanism of skin formation due to disulphide cross linking at high temperatures and concentrations. Powder properties were also established for selected products produced on the commercial scale driers. These powders were then used in experiments on the two smaller driers. Because collision frequency depends on the velocity and droplet size of sprays; these properties were measured for the small scale drier and estimated, where possible, for the pilot and commercial driers. The small scale agglomerating spray drier was configured to alter droplet and particle properties when interacting a vertical fines particle curtain with a horizontal spray sheet. An extensive design and improvement process was carried out to ensure the system consistently delivered these streams in a controllable manner. The processes of collision and adhesion occur very quickly inside the spray drier. In order to assess the extent of agglomeration that has occurred, the feed streams must be compared to the final product stream. An ideal way to do this is to use an agglomeration index which compares the particle size distributions of the feed (fines recycle and spray streams) and the particle size distribution of the product stream (the agglomerated powder). The index described changes between these steams across the particle size distribution and is called an agglomeration efficiency, ξg. However, it was found that the presence of fines in the product of the one-pass design obscured the agglomerates formed. The agglomeration efficiency, ξg, was modified to become ξh which subtracted the fines stream from the agglomerated product distribution. In this way ξh models industrial operation where the fines are recycled, by effectively just comparing the spray and product streams entering and leaving the process. The small scale drier was used for an experimental study on natural and forced agglomeration, where the drier was operated with spray only, then with spray and fines. For natural agglomeration, SEM images of the product powder indicated that little agglomeration occurred between spray droplets. The product yield was unacceptably low (~ 40%) due to adhesion of spray droplets to the drying chamber wall opposing the horizontal spray. When the fines curtain was introduced in the forced agglomeration experiments, product yield increased above 50% because the fines acted as collectors for the spray droplets. However, the agglomeration performance of the modified spray drier was lower than expected. The equipment design was then optimised by considering three key issues; fines dispersion, droplet dispersion and stickiness, and agglomerate breakdown. Final experiments studied agglomeration at low fines to spray mass flux ratios and showed that increasing the fines size had a positive effect on agglomeration efficiency,ξh. The agglomeration study at pilot scale identified the effect of key variables, total solids, concentrate and fines flow rate, and fines size on the agglomeration efficiency. A dimensionless flux approach was used to explain the experimental results. The fines to spray mass flux ratio and the projected area flux ratio (at constant concentrate flow rate) were found to be the most suitable to represent the physical processes during agglomeration. Experimental results showed that a higher dimensionless flux resulted in more agglomeration and as well as small fines size and atomising low solids concentrate. The critical Stokes number highlighted the importance of particle size and collision velocity on the outcome of the collision as well as the importance of stickiness on adherence following the collision. A statistical analysis established a relational model for predicting the agglomeration efficiency based on fines size, total solids and the fines to spray mass flux ratio. This thesis has gained insight into agglomeration processes during spray drying and knowledge about how to define the extent of agglomeration. Practical findings from this research can have a significant impact on successful spray drying operation for instant powders. There are some practical steps to be taken industrially to promote the control of agglomerating spray driers. The first step is to measure and control the flow of fines recycled to the top of the spray drier. The next step is to validate the findings at industrial scale and link the agglomeration index to the bulk powder properties. However, there are many challenges that remain to be tackled in the area of milk powder agglomeration. Milk powder agglomeration at the top of the spray drier is a complex process involving many different variables. A more detailed study of the micro processes that occur during agglomeration will give increased understanding of the relationships between key operating variables and agglomerate properties.

Main purpose of this study was to increase protein expression in Chinese hamster ovary (CHO) cells by improving protein secretion of translated proteins. The goal was to find signal peptides from the screening of signal peptide libraries for improvement of protein secretion using a CHO-cell express selection system. Biopharmaceutical products, proteins such as monoclonal antibodies (mAbs), are most commonly produced using mammalian expression systems such as the expression in CHO cells. The posttranslational modifications of the proteins being expressed in CHO cells are similar to the expressional modifications in human cells, why the CHO cells are suitable for production of proteins used for human therapy. The expression of proteins in the cell is a complex mechanism, fundamentally depending on the DNA sequences in the cell nucleus. Secretion of translated proteins has been showed to be a bottleneck when improving expression. Secretion is initiated by the signal peptide, a n-terminal prolongation of the protein that is recognized by a signal recognition particle (SRP) when being translated by the ribosome. The sequence and structure of the signal peptide has been proved to affect secretion and altering the signal peptide could improve secretion even when changing signal peptide between different species. Designing variants of the signal peptides and analyzing protein expression might lead to improvements of the construct design and more protein produced from the cells, which would save time, money and material for the producer. To construct plasmids containing the gene of interest (GOI) and different signal peptides, several gene cloning methods were used. The plasmids were amplified using Escherichia coli (E. coli) transformation. The constructs were expressed by transfection into the CHO cell genome, and expression were analyzed using flow cytometry. When analyzing expression of a Fc-fusion protein with 5 different signal peptides, the signal peptide Azurocidin is the one showing highest expression levels in this study. In addition, IgG kapa and Albumin signal peptides did not show as high protein expression levels, even if they were better than the L1d and H5b signal peptides. Since signal peptides are exchangeable between proteins and species, it might be that Azurocidin is improving secretion and protein expression with other proteins than Fc-fusion proteins which would be an interesting aspect for further studies. When altering signal peptides with library sequences, the experimental challenges were crucial for the protein expression results and due to these issues, no library sequence could be seen to conquer others when it comes to protein expression levels. Transfection and cultivation procedures needs to be studied and improved before being able to draw conclusions about which signal peptide library sequences that might improve secretion and increase the protein expression.

This Master Thesis was conducted at Octapharma AB in Stockholm. Anticomplementary activity (ACA) is a measure of the product’s abilities to activate the complement system. IgG aggregates are mainly responsible for this activation. Two different performances of a method for determination of ACA in Octagam® are available. The two performances are based on the reference method for test of ACA in immunoglobulins in the European Pharmacopoeia Commission Guideline 6.0 (chapter 2.6.17). The method is carried out either in test tubes or on microtiter plates. The test tube method can be performed either in a manual manner or modified, being more automated. The latter performance has been applied in this study. The plate method is more automated than both of the tube methods. The plate method and the manual tube method have earlier seemed to result in different outcomes, which was the basis for this thesis. The plate method and the modified test tube method have been compared and robustness parameters have been studied in order to see which factors influence on the end result. The adequacy of using Human Biological Reference Preparation (human BRP) as a control for the ACA method in general has also been investigated. Samples of the product are outside the scope of this thesis and have not been investigated. According to this study, the plate method and the modified tube method are not comparable with regard to complement titration results and to ACA of the BRP control. A higher precision is gained with the plate method. This in combination with the higher degree of automation makes the plate method advantageous in several aspects. When it comes to the robustness of the ACA method in general, the sheep red blood cells (SRBC) used are critical. Haemolysin dilution and complement activity seem to be critical as well. Human BRP is, according to this study more adequate as a reference for the plate method than for the tube method. An In house control is believed to be more representative to the ACA method in general as it is of the same nature as the samples analysed, in contrast to the human BRP.

Virtually all extracellular proteins in humans are glycoproteins and likewise are many biopharmaceuticals. The glycosylation is directly correlated to biological function and stability of these proteins. The ability to isolate glycoproteins is thus of great importance in many applications. The most common isolation method for glycoproteins is affinity chromatography using lectins, a ubiquitous and versatile group of carbohydrate-binding proteins. The lectin Concanavalin A (ConA) has long been used for this purpose but suffers from undesired leakage into the eluate, causing an inquiry of alternative chromatography ligands or optimization of the ConA resin.In this study, a total of 20 different lectins, including ConA, were evaluated and compared in terms of suitability as ligands in affinity chromatography for glycoprotein isolation. The lectins’ binding to glycoproteins were studied, mainly through microtiter plate binding assays using a monoclonal IgG1 antibody and Conalbumin (Ovotransferrin). Further, sugar-specificities and potential eluting sugars for the lectins were examined through inhibition with eight different carbohydrates. Additionally, the glycoprotein binding and leakage of ConA columns were examined, and a potential leakagereducing treatment of ConA resin evaluated.ConA was found to be superior in binding to the investigated glycoproteins but exhibited a limited binding when immobilized to an agarose resin. This discrepancy is likely a consequence of structurally hidden glycans on the used glycoproteins and requirements of long residence time when used in a chromatographic setting. Binding competition with several sugars were investigated with a similar microtiter plate binding assay. This method displayed potential to predict the behaviour of sugars and their suitability as eluting agents in a chromatography column. The best eluting sugar for ConA was showed to be methylmannoside, ideally in combination with methylglucoside. Lastly, evaluation of ConA columns with a crosslinking glutaraldehyde-treatment showed that the ConA ligand leakage may be significantly reduced, although further studies and optimizations are needed.This study thus presents a repertoire of lectins and their differences in terms of glycoprotein-binding and sugar-specificity, as well as evaluations of ConA columns’ efficiency and potential leakage-prevention.