While protein therapeutics are indispensable in the treatment of a variety of diseases, including cancer, rheumatoid arthritis, and diabetes, key limitations including short half-lives, high immunogenicity, protein instability, and centralized production complicate long-term use and on-demand production. Site-specific polymer conjugation provides a method for mitigating these challenges while minimizing negative impacts on protein activity. However, the location-dependent effects of polymer conjugation are not well understood. Cell-free protein synthesis provides direct access to the synthesis environment and rapid synthesis times, enabling rapid evaluation of multiple conjugation sites on a target protein. Here, work is presented towards developing cell-free protein synthesis as a platform for both design and on-demand production of next generation polymer-protein therapeutics, including (1) eliminating endotoxin contamination in cell-free reagents for simplified therapeutic preparation, (2) improving shelf-stability of cell-free reagents via lyophilization for on-demand production, (3) coupling coarse-grain simulation with high-throughput cell-free protein synthesis to enable rapid identification of optimal polymer conjugation sites, and (4) optimizing cell-free protein synthesis for production of therapeutic proteins

<div><div><div><p>To prolong their long-term stability, protein molecules are commonly dispensed as lyophilized powders to be reconstituted before use. Evaluating the stability of these biomolecules in the solid state is routinely done by using various analytical techniques such as glass transition temperature, residual moisture content and other spectroscopic techniques. However, these techniques often show poor correlation with long term storage stability studies. As a result, time intensive long term storage stability studies are still the golden standard for evaluating protein formulations in the solid state. Over the past few years, our lab has developed solid-state hydrogen deuterium exchange- mass spectrometry (ssHDX-MS) as an analytical tool that probes the backbone of a protein molecule in the solid state. ssHDX-MS gives a snapshot of protein-matrix interactions in the solid state and has a quick turnaround of a few weeks as opposed to a few months for accelerated stability testing. Additionally, various studies in the past have demonstrated that ssHDX-MS can be used for a wide range of biomolecules and shows strong correlation to long term stability studies routinely employed.</p><p>The main aim of this dissertation is to provide an initial understanding of the mechanism behind ssHDX-MS in structured protein formulations. Specifically, this dissertation is an attempt at studying the effects of various experimental variables on the ssHDX-MS of myoglobin formulations as well as demonstrating the utility of this analytical technique. Firstly, the effects of varying temperature and relative humidity on ssHDX-MS of myoglobin formulations is studied with the help of statistical modeling. Secondly, the effects of pressure on ssHDX-MS of myoglobin formulations are evaluated at an intact and peptide digest levels. Finally, ssHDX-MS is used as a characterization tool to evaluate the effects of two different lyophilization methods on the structure and stability of myoglobin formulations. The results of studies described in this dissertation show ssHDX-MS to be sensitive to changes in experimental parameters, namely temperature, relative humidity, pressure, and excipients. Additionally, ssHDX-MS results were in good agreement with other routinely employed analytical and stability testing techniques when used to compare the effects of two lyophilization methods on myoglobin formulations.</p></div></div></div>

The thesis deals with the extraction of antioxidants from elder flowers and the way of storage obtained extract. The elder flowers which are obtained by cutting trees as a waste product and after their processing it could be used as an ingredient to cosmetics products for body and face skin or in a spa like additive to baths. In theoretical part are information about black elder, its botanical characteristics, utilization and about substances contained in it. There is also a chapter about lyophilization as a method which was used for concentration of extract. The last chapter describes theoretical information about liquid chromatography because this method was used for analysis of contents of extract. Experimental part describes optimalization steps in preparation of extract. It was found that the biggest content of polyphenolic compounds was when the proportion between flower and water was 1:10, optimal temperature of water for extraction is 100 °C and it was also found that the time of extraction don´t have any influence on content of polyphenols. Thanks to the lyophilization it was obtained dry light brown powder which is stable for long time. The lyophilizate was tested for some physical properties like pH, refractive index, solubility etc. There was also determined antioxidant activity by DPPH method and there was found quenching activity is 64,9 %. In determining of heavy metals in sample was found trace amounts of lead and chrome. By liquid chromatography was determined content of polyphenols, caffeic acid was in an amount 59,6 mgl1, chlorogenic acid 398 mgl1 and ferulic acid wasn´t detected. All of analysis was for 1% solution of extract because to cosmetic it could be just this amount to addition. In the last step was prepared a skin lotion with elder flower extract and it was tested for stability also was prepared an ointment from pork lard.

The Diploma thesis deals with use of selected probiotic strains Lactobacillus acidophilus and Bifidobacterium breve in different forms in food for infants. The theoretical part is focused on describing probiotics, encapsulation methods and intestinal gut microbiota of infants. Further, characterization of individual periods of infant feeding and food for infants were introduced. In experimental part the possibilities of encapsulation and lyophilisation of probiotic cells were observed. Probiotic cells were encapsulated into alginate particles. The encapsulator was used for preparation of particles and the most appropriate particles were prepared by encapsulation nozzle with size of 300 µm. Moreover, probiotics viability was monitored by Flow Cytometry, Fluorescence Microscopy and by cultivation (CFU method). Viability of probiotics was monitored during long-term storage in selected food for infants. The appropriate shelf life of non-lyophilized alginate particles in real food have been set at 1 to 2 months. Lyophilized alginate particles could be stored for more than 3 months. Finally, the stability of the particles and viability of encapsulated and non-encapsulated cells in the gastrointestinal tract conditions were also examined. The viabilities of lyophilized cells and cells encapsulated in lyophilized particles were also compared. From the results obtained, non-encapsulated probiotic bacteria cells are more susceptible to negative effects of digestive juices, the percentage of dead probiotic cells after digestion was approximately 80 %. On the other hand, alginate particles showed cell protection from digestive juices, after incomplete cell releasing from particles the percentage of dead probiotic cells did not exceed 20 %. After adequate rehydration, similar results were gained with lyophilized alginate particles. Lyophilized alginate particles have been determined to be the most suitable application form for infants’ food.

Many proteins are unstable in solution and must be formulated in the solid state. This has led to an increase in the use of lyophilized dosage forms. Lyophilization is a complicated processing method consisting of three major steps: freezing, primary drying, and secondary drying. This can lead to several formulation stability challenges including changes in ionization within the matrix, phase separation of the protein drug from added stabilizers, sufficient mobility within the system for movement of reactive species and protein side chains, and crystallization of excipients upon storage. Solid-State Nuclear Magnetic Resonance Spectroscopy (SSNMR) is used to characterize many important properties of lyophilized formulations including crystalline vs. amorphous content, polymorphic form, ionization profile, interaction between formulation components with domain sizes, and mobility within the cake matrix. In order to study ionization changes in lyophilized solids, SSNMR and UV/Vis Diffuse Reflectance spectroscopy were used. 13C-labeled fumaric, succinic, and butyric acids were added to formulations at various pH levels, and were used to quantify change in the ionization of the matrix by monitoring the ionization ratios of the carboxylic acid peaks using SSNMR. pH indicators were also added to the formulations and their ionization ratio was determined using UV/Visible Diffuse Reflectance Spectroscopy. The ionization profile in the solid state was compared with that in solution before lyophilization. A rank ordering of ionization shift was made in pharmaceutically relevant buffers. SSNMR proton relaxation times (1H T1 and 1H T1rho) for each formulation component can be compared to determine homogeneity within the lyophilized matrix. The concept of spin diffusion is used in order to determine the length scale on which the components are either homogeneous or phase separated. The domain size is typically 20-50 nm or 2-10 nm for 1H T1 and 1H T1rho, respectively. PVP and dextran polymers were phase separated on both domains for physical mixtures and lyophilized mixtures. BSA and lysozyme were both lyophilized with formulations containing sucrose, trehalose, or mannitol as the stabilizer. Mannitol crystallized, and the relaxation times showed phase separation. Sucrose and trehalose both formed homogeneous systems at both length scales when formulated in a 1:1 ratio with BSA or lysozyme. Aspartame was shown to be phase separated from trehalose. The SSNMR proton relaxation times were also used to measure the local mobility in the lyophilized matrix, as a timescale of picoseconds to nanoseconds is associated with the 1H T1 relaxation time. Mobility was monitored in formulations containing a fixed amount of sucrose and mannitol, but with a variable amount of an IgG2 protein. The 1H T1 relaxation times decreased as protein content increased. The formulations with the highest relaxation time (lowest mobility), was the most stable in accelerated temperature conditions as monitored by size exclusion chromatography and capillary isoelectric focusing. This method can be used to rank order the most stable formulations at time-zero. Anti-plasticization was also studied by formulating sorbitol in various ratios with trehalose. The 1H T1 relaxation times increased with increasing sorbitol content, while the glass transition temperature decreased. Sorbitol and trehalose glasses were also exposed to different temperature storage conditions. Sorbitol appears to promote aging, as the formulations with higher sorbitol content showed larger increases in proton relaxation time.

<div>Proteins are susceptible to physical and chemical degradation in solution, which can lead to the loss of therapeutic activity and increase the potential for immunogenic responses when administered. Many degradation reactions are mediated by water, and therefore the proteins are often formulated as solids in which degradation rates are slowed significantly. Lyophilization is the most common method for producing solid protein formulations, which removes the water by sublimation and desorption under vacuum from the frozen protein solutions. Lyophilization requires excipients to protect the protein from the inherent stresses involved in the process. Degradation can still occur during lyophilization and storage, and needs to be characterized in order to develop a successful formulation with desired storage stability. The analytical techniques to characterize solid-state proteins are limited, however, and many do not provide site-specific information and lack the ability to predict stability beforehand.</div><div>Recently, solid-state hydrogen-deuterium exchange mass spectrometry (ssHDX-MS) has been developed to characterize proteins in solid powders with peptide level resolution. The technique was found to be sensitive to formulation and process changes. The ssHDX-MS metrics are highly correlated to the long-term storage stability, suggesting that the method can serve as a formulation screening tool. This dissertation aims to evaluate the factors affecting ssHDX kinetics and to develop a mechanistic understanding of the exchange process in solid samples, which in turn will support the solid-state protein development and enable it to be conducted in a more a cost and time-effective way. First, the contribution of peptide-matrix interactions to deuterium incorporation kinetics in the absence of higher-order structure was assessed using lyophilized poly-D, L-alanine peptides. Deuterium incorporation depended on excipient type and D₂O_(g) activity in the solid samples. A reversible pseudo-first-order kinetic model was proposed and validated using the experimental data. Second, the reversibility of the hydrogen-deuterium exchange reaction in the solid-state was evaluated to support the ssHDX mechanistic model further. The reaction was found to be reversible irrespective of initial conditions and independent of the excipient type. Pre-hydration of the peptide samples prior to deuterium labeling did not affect deuterium incorporation in amorphous samples compared to the controls not subjected to pre-hydration. Third, the contribution of peptide secondary structure to deuterium uptake kinetics was quantified using structured PDLA analogs. The deuterium incorporation in structured peptides was less than that of the PDLA peptides suggesting that both peptide structure and peptide-matrix interactions contribute to ssHDX-MS. Finally, a quantitative data analysis method was presented that allows the interpretation of ssHDX-MS data of a protein relative to controls. Altogether, the findings present a comprehensive mechanistic understanding of the ssHDX-MS of proteins that is relevant to the industry.</div>

La leishmaniose viscérale est une maladie tropicale négligée et létale en l’absence de traitement. L’Amphotéricine B (AmB) est une molécule efficace mais sa forme conventionnelle, Fungizone®, conduit à une toxicité limitant les doses tandis que les formulations lipidiques moins toxiques telles que l’Ambisome® sont très coûteuses. Ainsi, le besoin de nouvelles formes thérapeutiques à la fois non toxiques et peu coûteuses subsiste actuellement. Dans ce travail, nous avons étudié deux solutions possibles : la Fungizone® chauffée (H-AmB) et une microémulsion chargée en AmB (MEAmB). En ce qui concerne la microémulsion, une forme lyophilisée est souhaitable afin de s’affranchir des risques d’hydrolyse et de contamination microbienne. Les objectifs de la thèse étaient de développer les deux nouvelles formes, d’évaluer la toxicité et l’efficacité de H-AmB et MEAmB contre Leishmania donovani (souche LV9) in-vitro et in-vivo et également d’optimiser la lyophilisation de la microémulsion.La microémulsion MEAmB est composée de gouttelettes sphériques dont le diamètre moyen est proche de 35 nm et a montré un comportement rhéologique de type newtonien. L’analyse spectroscopique de H-AmB a révélé la formation de super-agrégats qui sont moins toxiques que d’autres états d’agrégation. La MeAmB ainsi que l’H-AmB ont montré une activité antiparasitaire équivalente à celle d’AmBisome® sur les formes axénique et intramacrophagique de L. donovani. L’indice de sélectivité pour ces deux formulations est élevé, en contraste avec celui de la Fungizone® native. De plus, à la différence d’AmBisome®, elles ont montré une activité importante sur des souches résistantes à l’AmB. L’activité anti-leishmanienne in-vivo des nouvelles formulations est comparable aux formulations de référence. De même, aucune différence significative des marqueurs de toxicité rénale et hépatique n’a pu être observée. Ainsi, l’H-AmB et la MEAmB pourraient être considérées comme des traitements alternatifs de la leishmaniose viscérale, avec l’avantage d’être moins onéreuses à produire que l’Ambisome®.Afin d’optimiser le procédé de lyophilisation de la MEAmB, un plan d’expérience a été mis en œuvre. Ainsi, la taille des gouttelettes est minimisée par l’utilisation de 5% de maltose comme cryoprotectant, avec une température de congélation de -80°C et un temps total de lyophilisation de 24h. Par ailleurs, aucune modification significative de la teneur en AmB n’a été observée après la lyophilisation. Ainsi, la MEAmB lyophilisée est stable et pourrait éviter la dégradation due à la présence d’eau. / Visceral leishmaniasis is a neglected tropical disease that can be fatal if left untreated. Amphotericin B (AmB) is effective in the treatment of this disease, but the conventional formulation, Fungizone® has dose-limiting toxicity while the less toxic lipid-based forms such as Ambisome® are expensive. Therefore, the need for new therapeutic systems remains. In this respect, the heating of the Fungizone® formulation (H-AmB), and the development of a microemulsion (ME) containing AmB (MEAmB) are possible solutions. In addition, it is desirable to remove water from microemulsion systems in order to reduce instability due to microbiological contamination and hydrolysis. Therefore, the objective of this work was to develop and to evaluate the activity and toxicity in vitro and in vivo of H-AmB and MEAmB against Leishmania donovani (strain LV9) and, furthermore, to optimize a lyophilized microemulsion system containing AmB. Rheological, size and morphology studies showed that MEAmB presented average droplet sizes of 35 nm, a Newtonian behavior and spherical morphology. Spectroscopic characterization of H-AmB showed the formation of superaggregates, which are less toxic than the other states of aggregation. In-vitro evaluation on both the axenic and intramacrophagic amastigote forms showed that H-AmB and MEAmB showed similar activity to Ambisome®. A high selectivity index of H-AmB and MEAmB was observed compared with unheated Fungizone®. Furthermore, both new formulations showed high activity against AmB-resistant strains compared with Ambisome®. In-vivo experiments designed to evaluate their activity and toxicity did not reveal significant differences in activity between the new and reference formulations. There were no significant differences either in indicators of renal and hepatic toxicity. Therefore, both H-AmB and MEAmB can be used as an alternative for the treatment of LV9, presenting an advantage over Ambisome® in their lower costs of production. Therefore, a complete experimental design was performed in order to optimize the lyophilisation of the microemulsion system. It was observed that microemulsions with smaller droplet sizes were obtained using maltose as a cryoprotectant at a concentration of 5%, with freezing at -80 ° C, and a lyophilization process period of 24 h. Furthermore, it was observed that ME containing AmB showed no significant changes in drug content before and after the lyophilization process. Therefore, in its lyophilized form, the ME can remain stable and avoid degradation due to the presence of water.

<p>Lyophilization is the process of controllably removing the water content from a material with the objective of increasing its stability and, hence, its shelf life. This dissertation addresses two of the challenges faced by lyophilization, namely continuous temperature-monitoring and lengthy primary drying step.</p> <p>Continuous temperature monitoring of the product is imperative to a successful lyophilization process. It is more efficient to employ wireless temperature sensors rather than the conventional thermocouples. These wireless sensors need to keep a low profile that does not allow bulky battery attachment. Therefore, harvesting microwave energy is an excellent practice to power these sensors. Energy harvesting problem is twofold. One, designing an efficient flexible power-harvester (rectenna). To address this problem, we present a flexible rectenna with superior efficiency. While doing so, we establish the design procedure that can be followed for similar designs. Two, delivering sufficient power to the rectenna location inside the chamber. To address this problem, we propose two electromagnetic techniques, namely the statistical electromagnetics (SEM) and the electromagnetic time reversal (EMTR). These enable uniform power distribution and higher total efficiency.</p> As for the lengthy primary drying, to speed up the process, we propose RF-heating as a replacement for conventional heating. We establish a procedure for frequency selection based on the material under lyophilization and the geometrical properties of the freeze-drier’s chamber. The same techniques, SEM and EMTR are used. We conduct RF-assisted lyophilization processes based on SEM on different pharmaceutical bare excipients and on Myoglobin in four different excipients. The results confirm the superiority of the proposed technique in terms of drying time and heating uniformity.

Le corps humain est un ensemble de microflores où cohabitent bactéries, archées, virus et eucaryotes. Ces écosystèmes complexes sont appelés microbiotes. Parmi ceux-ci figure le microbiote intestinal qui compte 1011 à 1014 bactéries/g de selle. Les modifications de la flore intestinale peuvent être à l'origine de pathologies comme les diarrhées infectieuses. Il s'agit d'un véritable problème de santé publique puisqu'environ 2.16 millions de décès sont liés à cette pathologie chaque année. Les virus intestinaux jouent un rôle prépondérant mais les infections bactériennes restent également importantes. Le diagnostic de ces infections bactériennes reste compliqué puisque le microbiote intestinal comporte 75% d'espèces non cultivables. De plus, on ne dispose pas réellement d'une liste exhaustive des bactéries pouvant être responsables de diarrhées infectieuses. Nous avons donc choisi d'étudier le microbiote intestinal dans des selles normales et pathologiques, par une approche polyphasique alliant une étape préliminaire de concentration des selles diarrhéiques par la lyophilisation, à des techniques de culture et des méthodes de biologie moléculaire. Pour cela nous avons mis au point une nouvelle technologie pour la détection des entéropathogènes par hybridation sur puce à ADN permettant la détection des bactéries et des virus ADN entéropathogènes, en présence d'un témoin archae. Notre outil permet le diagnostic multiplexe des diarrhées infectieuses puisque nous avons correctement identifié un adénovirus et la bactérie Campylobacter jejuni présents dans une même selle. / The human body is a collection of microflora where cohabit bacteria, archaea, viruses and eukaryotes. These complex ecosystems are called microbiota. Among these is the intestinal microbiota that counts 1011 to 1014 bacteria/g of stool. Changes in the intestinal flora can cause of pathologies such as infectious diarrhea. This is a real public health problem since about 2.16 million deaths are related to this disease each year. Enteric viruses play a preponderant role but bacterial infections are also important. The diagnosis of bacterial infections is complicated because the intestinal microbiota includes 75% non-cultivable species. In addition, there is not really a list of bacteria could be responsible for infectious diarrhea. We therefore decided to study the intestinal microbiota in normal stool and also pathological stools by a polyphasic approach combining a preliminary step of diarrheal stools concentration by lyophilization, with cultivation techniques and molecular biology methods. We developed a new technology for the detection of enteropathogens by hybridization on DNA microarray for the detection of bacteria and enteric viruses (DNA) in the presence of a control archaea. Our tool allows multiplexed diagnostic of infectious diarrhea since we correctly identified an adenovirus and Campylobacter jejuni present in a same sample. This is the first DNA microarray for multiplex detection of bacteria and viruses (DNA) enteropathogens. An improvement of our protocol for nucleic acid extraction is proposed to allow the detection of RNA viruses such as rotavirus and calicivirus which are currently dominant.

Foi avaliado o fornecimento do colostro bovino liofilizado como alternativa de fonte inicial de imunoglobulinas para caprinos recém-nascidos bem como as características histológicas, considerando avaliações histomorfométricas, estereológicas, imunohistoquímicas e o número de células caliciformes no epitélio intestinal destes animais. Foram utilizados dois grupos de 15 cabritos, que receberam 5% do peso vivo de colostro bovino liofilizado (CBL) ou colostro caprino (CC), com 55 mg/mL de imunoglobulina G, às 0, 7 e 14 horas de vida. Amostras do duodeno, jejuno médio e íleo foram coletadas às 18, 36 e 96 horas de vida para análises histológica e imunohistoquímica. Três animais foram sacrificados logo após o nascimento, constituindo o grupo zero hora. O delineamento experimental foi inteiramente casualizado, sendo as variáveis morfométricas consideradas em arranjo fatorial 2x3+1, tendo como efeitos principais os dois tipos de colostro, os três horários de abate e o grupo zero hora. A concentração de IGF-I no colostro bovino liofilizado e colostro caprino foi de 158,71 e 356,32 ng/mL, respectivamente. A altura das vilosidades no jejuno mostrou-se superior (P<0,05), apresentando vilosidades 30,7% e 24,2% mais altas do que o duodeno e o íleo às 36h, respectivamente. O duodeno, às 18h, apresentou profundidade das criptas 48,4% maior que o jejuno e, às 96h as criptas duodenais mostraram-se 23,2% maiores que as criptas do íleo (P<0,05). No jejuno, as criptas apresentaram-se mais profundas às 96h (P<0,05) do que nos outros horários. A espessura da túnica muscular no jejuno apresentou-se mais espessa às 36 e 96h do que à zero h e mais espessa às 36h do que às 18h (P<0,05). No geral, jejuno e íleo, apresentaram túnica muscular menos espessa (P<0,05) que a encontrada no duodeno. Independente dos segmentos, às 96h a túnica muscular foi mais espessa que às 18h. O Vv da mucosa absortiva foi maior no jejuno, enquanto no íleo às 36h o Vv foi maior do que nos demais horários experimentais. Independente do segmento, às 36h o Vv foi 17,3% maior que às 96h. O número de células caliciformes no jejuno foi 78,6% menor às 18h (P<0,05) nos cabritos que ingeriram colostro caprino e 54,6% maior às 96h (P<0,05) nos animais que ingeriram colostro bovino liofilizado. Colostro bovino liofilizado pode ser utilizado como fonte de proteção passiva alternativa para os caprinos recémnascidos. O IGF-I presente no colostro não influenciou a morfometria entérica. O jejuno mostrou-se o segmento mais importante no processo absortivo de imunoglobulinas. A distribuição dos anticorpos internalizados pelo epitélio entérico mostrou-se relacionada aos segmentos do intestino e tempo de vida dos recém-nascidos, independente da fonte de anticorpos, bovina liofilizada ou caprina. / It was evaluated the supply of the lyophilized bovine colostrum as an alternative source of immunoglobulin for newborn goat kids and the histological characteristics, through histomorphometric, stereological, immunohistochemical evaluations and goblet cells number in the intestinal epithelium in this animals. Two groups of fifteen newborn goat kids received 5% of body weight of lyophilized bovine colostrum (LBC) or caprine colostrum (CC) with 55 mg/mL of immunoglobulin G at 0, 7 and 14 hr of life. Samples of duodenum, medium jejunum and ileum were collected at 18, 36 and 96 hr of life for histological and immunohistochemical analysis. Three animals were sampled just after birth representing the zero hr group. A completely randomized designing was used, and the variable morphometric was considered as 2x3+1 factorial arrangement, having as the main factors colostrum supply, the three slaughter hours and the zero hr group. The IGF-I concentration in lyophilized bovine colostrum or caprine colostrum was 158,71 and 356,32 ng/mL, respectively. Villous height was superior in the jejunum (P<0.05) with villous 30.7% and 24.2% higher than duodenum and ileum at 36hr, respectively. The duodenum showed at 18hr crypts 48.4% deeper than in the jejunum and at 96hr, the duodenal crypts was 23.2% deeper than ileum crypts (P<0.05). In the jejunum, crypts were deeper at 96hr compared to the other experimental hours. The muscularis submucosal thickness in the jejunum was thicker at 36 and 96hr than zero hr and was thicker at 36hr than 18hr. Jejunum and ileum, in general thinner (P<0.05) tunica muscular layer than found in the duodenum. Regardless of the segments, at 96hr, the muscularis submucosal was thicker than 18hr. The absorptive mucosa partial volume (Vv) was higher in the jejunum, while in the ileum the Vv was higher at 36hr than all other experimental dates. Regardless of the segments, at 36hr, the Vv was 17.3% higher than at 96hr. The goblet cells number was different in jejunum with 78.6% less cells at 18hr (P<0.05) in the goat kids fed caprine colostrum and 54.6% higher at 96hr (P<0.05) in the goat kids fed lyophilized colostrum. Lyophilized bovine colostrum can be used as an alternative source of passive protection for the newborn kids. The IGF-I present in colostrum didnt influence the enteric morphology. The jejunal epithelium was the most important segment related to absorption process. The internalized antibody distribution in the enteric epithelium showed related to the small intestine segments and post-partum period, regardless of the antibodies source, lyophilized bovine or caprine.