Capillary zone electrophoresis (CZE) is a widely used analytical method. CZE is described theoretically very well and there are many simulation programs, which enable one to predict results of electrophoretic separations, and alternatively to study phenomena taking place during the electrophoretic separation in detail. The CZE method is not only an analytical method, but is often used for determination of many physical parameters of compounds, such as stability constants or complex mobilities. Among methods most often used for determination of complexation parameters belongs the affinity capillary electrophoresis (ACE). Its alternative is the vacancy affinity capillary electrophoresis (VACE). Whereas by the ACE method the stability constant is determined from the dependence of the analyte effiective electrophoretic mobility on the background electrolyte (BGE) composition, in the case of the VACE system peaks are used for this purpose. In this work the legitimacy of using system peaks in the VACE method for determination of stability constants was investigated. Several approaches dealing with the concentrating of complexing agent in the peak area were compared, both for the ACE and the VACE method. Two different kinds of electrophoretic systems were studied. In the first one, neutral cyclodextrin was used as...

Dans notre thèse, nous étudions les rapports que les professeurs construisent avec les disciplines qu’ils enseignent en proposant deux concepts : l’affinité disciplinaire et l’affinité didactique. Nous nous intéressons plus particulièrement aux enseignants et futurs enseignants de sciences physiques et chimiques à la fois du point de vue des fondements et des conditions d’émergence des affinités disciplinaire et didactique, ainsi que leurs effets sur l’enseignement, notamment dans le contexte d’un nouveau programme de lycée, le travail documentaire et les interactions avec les collègues. Nous avons mobilisé deux cadres théoriques : l’analyse praxéologique (Chevallard, 1998) et l’approche documentaire du didactique (Gueudet & Trouche, 2008). Nous avons combiné différents outils de recueil de données : questionnaires, entretiens, observations de classe, et visites des systèmes de ressources des enseignants. Les trois derniers outils ont été appliqués à un petit nombre de professeurs, choisis en fonction de leur affinité déclarée et de leur place dans le processus de constitution des ressources d’enseignement (étudiants en master se destinant à l’enseignement et enseignants expérimentés). Nous avons ciblé l’enseignement d’un concept précis, le spectre, choisi pour sa position frontière (issu de la physique et exploité en chimie par les méthodes spectroscopiques). La thèse propose des développements théoriques et méthodologiques pour saisir les interactions entre les affinités disciplinaire et didactique des enseignants et leur travail documentaire. Les résultats montrent l’existence d’une affinité pour une des deux disciplines, et elle met en évidence les relations fortes entre genèse de l’affinité didactique et genèse des ressources de l’enseignant. / In our thesis, we study the relationships that teachers build with the disciplines that they teach by offering two concepts: disciplinary affinity and didactic affinity.We focus on physics and chemistry, both in terms of the basis and conditions for the emergence of disciplinary and didactic affinities, their effects on teaching, particularly in the context of a new curriculum, documentary work and interactions with colleagues. We have mobilized two theoretical frameworks: the anthropological approach of didactics (Chevallard, 1998) and the documentary approach to didactics (Gueudet & Trouche, 2008). We combined several methodological tools: questionnaries, interviews, classroom observations, and teachers resource systems visits. The last three tools have been applied to a small number of teachers selected with respect to their claimed affinity and their place in the process of teaching resources design (pre-service vs. experienced teachers) targeting the teaching of a specific concept, the spectrum, chosen for its border position (from physics and chemistry operated by spectroscopic methods).The thesis offers theoretical and methodological developments to capture interactions between disciplinary and didactic affinities of teachers and their documentary work. The results evidence the presence of an affinity for one of the two disciplines, and it highlights the strong relationship between genesis of didactic affinity and genesis of teaching resources.

Amid the 2011 Arab Revolts, and the subsequent worldwide Occupy movement, social movement scholars faced sudden, powerful mass mobilisations without easily identifiable resources, networks, or forms of organisation underlying them. These instances of mobilisation beyond the scope of what we traditionally consider ‘the movement’ have stretched existing theories of social movements to their limits, defying both conventional theoretical frameworks and existing approaches. This work undertakes a novel analysis of mobilisation which accounts for these new, disruptive cases. It advances the concept of Affinity: a predisposition to participate in certain causes based on social or psychological traits. Alongside this concept, it outlines conditions of Convergence: emergent situations, frames and spaces which encourage those with such Affinity to temporarily participate in mass mobilisations. These two concepts are advanced and developed through a study of the 2011 Egyptian Revolt and Occupy Wall Street movement, alongside the classic case of the 1789 French Revolution. These cases are analysed in comparative perspective to develop a powerful analytical tool with which scholars can augment conventional analyses: The Affinity-Convergence Model of Mobilisation.

Recombinant strategies are today dominating in thedevelopment of modern subunit vaccines. This thesis describesstrategies for the production and recovery of protein subunitimmunogens, and how genetic design of the expression vectorscan be used to adapt the immunogens for incorporation intoadjuvant systems. In addition, different strategies fordelivery of subunit vaccines by RNA or DNA immunization havebeen investigated. Attempts to create general production strategies forrecombinant protein immunogens in such a way that these areadapted for association with an adjuvant formulation wereevaluated. Different hydrophobic amino acid sequences, beingeither theoretically designed or representing transmembraneregions of bacterial or viral origin, were fused on gene leveleither N-terminally or C-terminally to allow association withiscoms. In addition, affinity tags derived fromStaphylococcus aureusprotein A (SpA) or streptococcalprotein G (SpG), were incorporated to allow efficient recoveryby means of affinity chromatography. A malaria peptide, M5,derived from the central repeat region of thePlasmodium falciparumblood-stage antigen Pf155/RESA,served as model immunogen in these studies. Furthermore,strategies forin vivoorin vitrolipidation of recombinant immunogens for iscomincorporation were also investigated, with a model immunogendeltaSAG1 derived fromToxoplasma gondii. Both strategies were found to befunctional in that the produced and affinity purified fusionproteins indeed associated with iscoms. The iscoms werefurthermore capable of inducing antigen-specific antibodyresponses upon immunization of mice, and we thus believe thatthe presented strategies offer convenient methods for adjuvantassociation. Recombinant production of a respiratory syncytial virus(RSV) candidate vaccine, BBG2Na, in baby hamster kidney(BHK-21) cells was investigated. Semliki Forest virus(SFV)-based expression vectors encoding both intracellular andsecreted forms of BBG2Na were constructed and found to befunctional. Efficient recovery of BBG2Na could be achieved bycombining serum-free production with a recovery strategy usinga product-specific affinity-column based on a combinatoriallyengineered SpA domain, with specific binding to the G proteinpart of the product. Plasmid vectors encoding cytoplasmic or secreted variants ofBBG2Na, and employing the SFV replicase for self-amplification,was constructed and evaluated for DNA immunization against RSV.Both plasmid vectors were found to be functional in terms ofBBG2Na expression and localization. Upon intramuscularimmunization of mice, the plasmid vector encoding the secretedvariant of the antigen elicited significant anti-BBG2Na titersand demonstrated lung protective efficacy in mice. This studyclearly demonstrate that protective immune responses to RSV canbe elicited in mice by DNA immunization, and that differentialtargeting of the antigens expressed by nucleic acid vaccinationcould significantly influence the immunogenicity and protectiveefficacy. We further evaluated DNA and RNA constructs based on the SFVreplicon in comparison with a conventional DNA plasmid forinduction of antibody responses against theP. falciparumPf332-derived antigen EB200. In general,the antibody responses induced were relatively low, the highestresponses surprisingly obtained with the conventional DNAplasmid. Also recombinant SFV suicide particles inducedEB200-reactive antibodies. Importantly, all immunogens inducedan immunological memory, which could be efficiently activatedby a booster injection with EB200 protein. <b>Keywords</b>: Affibody, Affinity chromatography, Affinitypurification, DNA immunization, Expression plasmid, Fusionprotein, Hydrophobic tag, Iscoms, Lipid tagging, Malaria,Mammalian cell expression, Recombinant immunogen, RespiratorySyncytial Virus, Semliki Forest virus, Serum albumin,Staphylococcus aureusprotein A, Subunit vaccine,Toxoplasma gondii

In order to produce predictable and robust systems forprotein purification and detection, well characterized, small,folded domains descending from bacterial receptors have beenused. These bacterial receptors, staphylococcal protein A (SPA)and streptococcal protein G (SPG), possess high affinity to IgGand / or HSA. They are composed of repetitive units in whicheach one binds the ligand independently. The domains foldindependently and are very stable. Since the domains also havewellknown three-dimensional structures and do not containcysteine residues, they are very suitable as frameworks forfurther protein engineering. Streptococcal protein G (SPG) is a multidomain proteinpresent on the cell surface ofStreptococcus. X-ray crystallography has been used todetermine the binding site of the Ig-binding domain. In thisthesis the region responsible for the HSA affinity of ABD3 hasbeen determined by directed mutagenesis followed by functionaland structural analysis. The analysis shows that the HSAbindinginvolves residues mainly in the second α-helix. Most protein-based affinity chromatography media are verysensitive towards alkaline treatment, which is the preferredmethod for regeneration and removal of contaminants from thepurification devices in industrial applications. Here, aprotein engineering strategy has been used to improve thetolerance to alkaline conditions of different domains fromprotein G, ABD3 and C2. Amino acids known to be susceptibletowards high pH were substituted for less alkali susceptibleresidues. The new, engineered variants of C2 and ABD shownhigher stability towards alkaline pH. Also, very important forthe potential use as affinity ligands, these mutated variantsretained the secondary structure and the affinity to HSA andIgG, respectively. Moreover, dimerization was performed toinvestigate whether a higher binding capacity could be obtainedby multivalency. For ABD, binding studies showed that divalentligands coupled using non-directed chemistry demonstrated anincreased molar binding capacity compared to monovalentligands. In contrast, equal molar binding capacities wereobserved for both types of ligands when using a directed ligandcoupling chemistry involving the introduction and recruitmentof a unique C-terminal cysteine residue. The staphylococcal protein A-derived domain Z is also a wellknown and thoroughly characterized fusion partner widely usedin affinity chromatography systems. This domain is consideredto be relatively tolerant towards alkaline conditions.Nevertheless, it is desirable to further improve the stabilityin order to enable an SPA-based affinity medium to withstandeven longer exposure to the harsh conditions associated withcleaning in place (CIP) procedures. For this purpose adifferent protein engineering strategy was employed. Smallchanges in stability due to the mutations would be difficult toassess. Hence, in order to enable detection of improvementsregarding the alkaline resistance of the Z domain, a by-passmutagenesis strategy was utilized, where a mutated structurallydestabilized variant, Z(F30A) was used as a surrogateframework. All eight asparagines in the domain were exchangedone-by-one. The residues were all shown to have differentimpact on the alkaline tolerance of the domain. By exchangingasparagine 23 for a threonine we were able to remarkablyincrease the stability of the Z(F30A)-domain towards alkalineconditions. Also, when grafting the N23T mutation to the Zscaffold we were able to detect an increased tolerance towardsalkaline treatment compared to the native Z molecule. In allcases, the most sensitive asparagines were found to be locatedin the loops region. In summary, the work presented in this thesis shows theusefulness of protein engineering strategies, both to explorethe importance of different amino acids regarding stability andfunctionality and to improve the characteristics of aprotein. <b>Keywords:</b>binding, affinity, human serum albumin (HSA),albumin-binding domain (ABD), affinity chromatography,deamidation, protein A, stabilization, Z-domain, capacity,protein G, cleaning-in-place (CIP), protein engineering, C2receptor.

Potassium (K+) is an essential nutrient and the most abundant cation in plant cells. Plants possess two transport systems for K+ acquisition: a high-affinity system (HATS), operating at external K+ concentrations ([K+]ext) below 1 mM, and showing reduced transport activity in the presence of ammonium (NH4+); and, a low-affinity system (LATS), operating at [K+]ext above 1 mM, that is not affected by NH4+. K+ transport and compartmentation were investigated in barley (Hordeum vulgare L.) and rice (Oryza sativa L.) using the non-invasive technique of compartmental analysis by tracer efflux (CATE), to simultaneously determine unidirectional membrane fluxes, ion concentrations, and exchange characteristics in subcellular compartments. These studies revealed striking differences in unidirectional K+ fluxes between HATS and LATS. It was found that flux measurements, using traditional direct influx (DI) protocols, accurately represented HATS influx, but underestimated LATS influx by as much as seven-fold. In both barley and rice, LATS K+ fluxes were found to undergo rapid, futile cycling, with the ratio of efflux:influx 3 to 5 times greater, and the cytosolic exchange rate 2 to 3 times faster than under HATS. Based upon plasma-membrane electrical potential measurements, efflux was found to be active under LATS conditions. LATS-mediated conditions for K+ were found to provide relief from NH4+ toxicity in barley by immediately reducing NH4+ influx by more than 50%, and significantly reducing NH4+ futile cycling. Employing the K+ channel inhibitors cesium, lanthanum, and tetraethylammonium, NH4+ was shown to have both K+-sensitive and –insensitive influx pathways at high [NH4+]ext. Based on current models of flux energetics, the combined uptake of K+ and NH4+ was found to utilize 60% of root oxygen consumption. Barley and rice both showed signs of NH4+ toxicity at low [K+]ext, but rice recovered at much lower [K+]ext, suggesting a crucial role of K+ in the NH4+-tolerance of rice. These experiments address fundamental aspects of K+ fluxes, and help provide a physiological framework for future studies of K+ transport and mineral nutrition.

Potassium (K+) is an essential nutrient and the most abundant cation in plant cells. Plants possess two transport systems for K+ acquisition: a high-affinity system (HATS), operating at external K+ concentrations ([K+]ext) below 1 mM, and showing reduced transport activity in the presence of ammonium (NH4+); and, a low-affinity system (LATS), operating at [K+]ext above 1 mM, that is not affected by NH4+. K+ transport and compartmentation were investigated in barley (Hordeum vulgare L.) and rice (Oryza sativa L.) using the non-invasive technique of compartmental analysis by tracer efflux (CATE), to simultaneously determine unidirectional membrane fluxes, ion concentrations, and exchange characteristics in subcellular compartments. These studies revealed striking differences in unidirectional K+ fluxes between HATS and LATS. It was found that flux measurements, using traditional direct influx (DI) protocols, accurately represented HATS influx, but underestimated LATS influx by as much as seven-fold. In both barley and rice, LATS K+ fluxes were found to undergo rapid, futile cycling, with the ratio of efflux:influx 3 to 5 times greater, and the cytosolic exchange rate 2 to 3 times faster than under HATS. Based upon plasma-membrane electrical potential measurements, efflux was found to be active under LATS conditions. LATS-mediated conditions for K+ were found to provide relief from NH4+ toxicity in barley by immediately reducing NH4+ influx by more than 50%, and significantly reducing NH4+ futile cycling. Employing the K+ channel inhibitors cesium, lanthanum, and tetraethylammonium, NH4+ was shown to have both K+-sensitive and –insensitive influx pathways at high [NH4+]ext. Based on current models of flux energetics, the combined uptake of K+ and NH4+ was found to utilize 60% of root oxygen consumption. Barley and rice both showed signs of NH4+ toxicity at low [K+]ext, but rice recovered at much lower [K+]ext, suggesting a crucial role of K+ in the NH4+-tolerance of rice. These experiments address fundamental aspects of K+ fluxes, and help provide a physiological framework for future studies of K+ transport and mineral nutrition.

Fragment-based drug discovery is an emerging process that has gained popularity in recent years. The process starts from small molecules called fragments. One major step in fragment-based drug discovery is fragment screening, which is a strategy to screen libraries of small molecules to find hits. The strategy in theory is more efficient than traditional high-throughput screening that works with larger molecules. As fragments intrinsically possess weak affinity to a target, detection techniques of high sensitivity to affinity are required for fragment screening. Furthermore, the use of different screening methods is necessary to improve the likelihood of success in finding suitable fragments. Since no single method can work for all types of screening, there is a demand for new techniques. The aim of this thesis is to introduce weak affinity chromatography (WAC) as a novel technique for fragment screening. WAC is, as the name suggests, an affinity-based liquid chromatographic technique that separates compounds based on their different weak affinities to an immobilized target. The higher affinity a compound has towards the target, the longer it remains in the separation unit, and this will be expressed as a longer retention time. The affinity measure and ranking of affinity can be achieved by processing the obtained retention times of analyzed compounds. In this thesis, WAC is studied for fragment screening on two platforms. The first system comprised a 24-channel affinity cartridge that works in cooperation with an eight-needle autosampler and 24 parallel UV detector units. The second system was a standard analytical LC-MS platform that is connected to an affinity column, generally called WAC-MS or affinity LC-MS. The evaluation criteria in studying WAC for fragment screening using these platforms were throughput, affinity determination and ranking, specificity, operational platform characteristics and consumption of target protein and sample. The model target proteins were bovine serum albumin for the first platform, thrombin and trypsin for the latter. Screened fragments were either small molecule drugs, a thrombin-directed collection of compounds, or a general-purpose fragment library. To evaluate WAC for early stages of fragment elaboration, diastereomeric mixtures from a thrombin-directed synthesis project were screened. Although both analytical platforms can be used for fragment screening, WAC-MS shows more useful features due to easy access to the screening platform, higher throughput and ability to analyze mixtures. Affinity data from WAC are in good correlation with IC50 values from enzyme assay experiments. The possibility to distinguish specific from non- specific interactions plays an important role in the interpretation of WAC results. In this thesis, this was achieved by inhibiting the active site of the target protein to measure off-site interactions. WAC proves to be a sensitive, robust, moderate in cost and easy to access technique for fragment screening, and can also be useful in the early stages of fragment evolution. In conclusion, this thesis has demonstrated the proof of principle of using WAC as a new tool to monitor affinity and to select hits in fragment-based drug discovery. This thesis has indicated the primary possibilities, advantages as well as the limitations of WAC in fragment screening procedures.  In the future, WAC should be evaluated on other targets and fragment libraries in order to realize more fully the potential of the technology.

<p>Recombinant strategies are today dominating in thedevelopment of modern subunit vaccines. This thesis describesstrategies for the production and recovery of protein subunitimmunogens, and how genetic design of the expression vectorscan be used to adapt the immunogens for incorporation intoadjuvant systems. In addition, different strategies fordelivery of subunit vaccines by RNA or DNA immunization havebeen investigated.</p><p>Attempts to create general production strategies forrecombinant protein immunogens in such a way that these areadapted for association with an adjuvant formulation wereevaluated. Different hydrophobic amino acid sequences, beingeither theoretically designed or representing transmembraneregions of bacterial or viral origin, were fused on gene leveleither N-terminally or C-terminally to allow association withiscoms. In addition, affinity tags derived from<i>Staphylococcus aureus</i>protein A (SpA) or streptococcalprotein G (SpG), were incorporated to allow efficient recoveryby means of affinity chromatography. A malaria peptide, M5,derived from the central repeat region of the<i>Plasmodium falciparum</i>blood-stage antigen Pf155/RESA,served as model immunogen in these studies. Furthermore,strategies for<i>in vivo</i>or<i>in vitro</i>lipidation of recombinant immunogens for iscomincorporation were also investigated, with a model immunogendeltaSAG1 derived from<i>Toxoplasma gondii</i>. Both strategies were found to befunctional in that the produced and affinity purified fusionproteins indeed associated with iscoms. The iscoms werefurthermore capable of inducing antigen-specific antibodyresponses upon immunization of mice, and we thus believe thatthe presented strategies offer convenient methods for adjuvantassociation.</p><p>Recombinant production of a respiratory syncytial virus(RSV) candidate vaccine, BBG2Na, in baby hamster kidney(BHK-21) cells was investigated. Semliki Forest virus(SFV)-based expression vectors encoding both intracellular andsecreted forms of BBG2Na were constructed and found to befunctional. Efficient recovery of BBG2Na could be achieved bycombining serum-free production with a recovery strategy usinga product-specific affinity-column based on a combinatoriallyengineered SpA domain, with specific binding to the G proteinpart of the product.</p><p>Plasmid vectors encoding cytoplasmic or secreted variants ofBBG2Na, and employing the SFV replicase for self-amplification,was constructed and evaluated for DNA immunization against RSV.Both plasmid vectors were found to be functional in terms ofBBG2Na expression and localization. Upon intramuscularimmunization of mice, the plasmid vector encoding the secretedvariant of the antigen elicited significant anti-BBG2Na titersand demonstrated lung protective efficacy in mice. This studyclearly demonstrate that protective immune responses to RSV canbe elicited in mice by DNA immunization, and that differentialtargeting of the antigens expressed by nucleic acid vaccinationcould significantly influence the immunogenicity and protectiveefficacy.</p><p>We further evaluated DNA and RNA constructs based on the SFVreplicon in comparison with a conventional DNA plasmid forinduction of antibody responses against the<i>P. falciparum</i>Pf332-derived antigen EB200. In general,the antibody responses induced were relatively low, the highestresponses surprisingly obtained with the conventional DNAplasmid. Also recombinant SFV suicide particles inducedEB200-reactive antibodies. Importantly, all immunogens inducedan immunological memory, which could be efficiently activatedby a booster injection with EB200 protein.</p><p><b>Keywords</b>: Affibody, Affinity chromatography, Affinitypurification, DNA immunization, Expression plasmid, Fusionprotein, Hydrophobic tag, Iscoms, Lipid tagging, Malaria,Mammalian cell expression, Recombinant immunogen, RespiratorySyncytial Virus, Semliki Forest virus, Serum albumin,<i>Staphylococcus aureus</i>protein A, Subunit vaccine,<i>Toxoplasma gondii</i></p>

<p>In order to produce predictable and robust systems forprotein purification and detection, well characterized, small,folded domains descending from bacterial receptors have beenused. These bacterial receptors, staphylococcal protein A (SPA)and streptococcal protein G (SPG), possess high affinity to IgGand / or HSA. They are composed of repetitive units in whicheach one binds the ligand independently. The domains foldindependently and are very stable. Since the domains also havewellknown three-dimensional structures and do not containcysteine residues, they are very suitable as frameworks forfurther protein engineering.</p><p>Streptococcal protein G (SPG) is a multidomain proteinpresent on the cell surface of<i>Streptococcus</i>. X-ray crystallography has been used todetermine the binding site of the Ig-binding domain. In thisthesis the region responsible for the HSA affinity of ABD3 hasbeen determined by directed mutagenesis followed by functionaland structural analysis. The analysis shows that the HSAbindinginvolves residues mainly in the second α-helix.</p><p>Most protein-based affinity chromatography media are verysensitive towards alkaline treatment, which is the preferredmethod for regeneration and removal of contaminants from thepurification devices in industrial applications. Here, aprotein engineering strategy has been used to improve thetolerance to alkaline conditions of different domains fromprotein G, ABD3 and C2. Amino acids known to be susceptibletowards high pH were substituted for less alkali susceptibleresidues. The new, engineered variants of C2 and ABD shownhigher stability towards alkaline pH. Also, very important forthe potential use as affinity ligands, these mutated variantsretained the secondary structure and the affinity to HSA andIgG, respectively. Moreover, dimerization was performed toinvestigate whether a higher binding capacity could be obtainedby multivalency. For ABD, binding studies showed that divalentligands coupled using non-directed chemistry demonstrated anincreased molar binding capacity compared to monovalentligands. In contrast, equal molar binding capacities wereobserved for both types of ligands when using a directed ligandcoupling chemistry involving the introduction and recruitmentof a unique C-terminal cysteine residue.</p><p>The staphylococcal protein A-derived domain Z is also a wellknown and thoroughly characterized fusion partner widely usedin affinity chromatography systems. This domain is consideredto be relatively tolerant towards alkaline conditions.Nevertheless, it is desirable to further improve the stabilityin order to enable an SPA-based affinity medium to withstandeven longer exposure to the harsh conditions associated withcleaning in place (CIP) procedures. For this purpose adifferent protein engineering strategy was employed. Smallchanges in stability due to the mutations would be difficult toassess. Hence, in order to enable detection of improvementsregarding the alkaline resistance of the Z domain, a by-passmutagenesis strategy was utilized, where a mutated structurallydestabilized variant, Z(F30A) was used as a surrogateframework. All eight asparagines in the domain were exchangedone-by-one. The residues were all shown to have differentimpact on the alkaline tolerance of the domain. By exchangingasparagine 23 for a threonine we were able to remarkablyincrease the stability of the Z(F30A)-domain towards alkalineconditions. Also, when grafting the N23T mutation to the Zscaffold we were able to detect an increased tolerance towardsalkaline treatment compared to the native Z molecule. In allcases, the most sensitive asparagines were found to be locatedin the loops region.</p><p>In summary, the work presented in this thesis shows theusefulness of protein engineering strategies, both to explorethe importance of different amino acids regarding stability andfunctionality and to improve the characteristics of aprotein.</p><p><b>Keywords:</b>binding, affinity, human serum albumin (HSA),albumin-binding domain (ABD), affinity chromatography,deamidation, protein A, stabilization, Z-domain, capacity,protein G, cleaning-in-place (CIP), protein engineering, C2receptor.</p>