Improving selection of alphaIIbbeta3-binding phage antibodies with increased reactivity derived from immunized donors

Although many studies of the immune response in polytransfused Glanzmann thrombasthenia (GT) patients and in autoimmune thrombocytopenic purpura (AITP) have demonstrated the frequent development of Abs directed against the alphaIIbbeta3 integrin, little is known about the induced anti-alphaIIbbeta3...

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Veröffentlicht in:Clinical immunology (Orlando, Fla.). - 1999. - 108(2003), 3 vom: 19. Sept., Seite 199-210
1. Verfasser: Jacobin, M J (VerfasserIn)
Weitere Verfasser: Robert, R, Pouns, O, Laroche-Traineau, J, Nurden, A, Peter, K, Little, M, Clofent-Sanchez, G
Format: Aufsatz
Sprache:English
Veröffentlicht: 2003
Zugriff auf das übergeordnete Werk:Clinical immunology (Orlando, Fla.)
Schlagworte:Case Reports Journal Article Research Support, Non-U.S. Gov't Antibodies, Monoclonal Autoantibodies Complementarity Determining Regions Immunoglobulin Heavy Chains Immunoglobulin Light Chains Oligopeptides Peptide Library mehr... Platelet Glycoprotein GPIIb-IIIa Complex arginyl-glycyl-aspartic acid 78VO7F77PN
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100 1 |a Jacobin, M J  |e verfasserin  |4 aut 
245 1 0 |a Improving selection of alphaIIbbeta3-binding phage antibodies with increased reactivity derived from immunized donors 
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500 |a Date Revised 08.11.2019 
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520 |a Although many studies of the immune response in polytransfused Glanzmann thrombasthenia (GT) patients and in autoimmune thrombocytopenic purpura (AITP) have demonstrated the frequent development of Abs directed against the alphaIIbbeta3 integrin, little is known about the induced anti-alphaIIbbeta3 autoantibodies at the molecular level. Phage display is a powerful technology for selecting and engineering mAbs expressed on the surface of filamentous bacteriophage. Combinatorial libraries of single-chain IgG were constructed from splenocytes from two patients with AITP and one patient with GT. In a previous study, activated platelets or alphaIIbbeta3-expressing CHO cells selection was performed to isolate human IgG anti-alphaIIbbeta3 binding fragments using combinatorial libraries created from the B cells of a GT and an AITP patient. However, we have experienced practical problems such as enrichment of truncated antibodies during selection. We decided to test prolonged treatments with elution agents after screening on the purified form of the alphaIIbbeta3 integrin activated with the RGD peptide. We obtained a higher percentage of clones with full-size antibody fragments as well as an enrichment of more specific alphaIIbbeta3-binding phage-Abs. Some of them, recognizing the activated form of the integrin, would be interesting to further study as potential diagnostic or therapeutic agents in acute coronary syndromes. Sequencing of selected phage-Abs revealed that they used different VH and VL genes with, for the majority of them, a high level of extensive hypermutations in the complementarity determining regions, indicating the diversity of the antigen-driven immune response that occurred in GT and AITP patients 
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650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
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650 7 |a Autoantibodies  |2 NLM 
650 7 |a Complementarity Determining Regions  |2 NLM 
650 7 |a Immunoglobulin Heavy Chains  |2 NLM 
650 7 |a Immunoglobulin Light Chains  |2 NLM 
650 7 |a Oligopeptides  |2 NLM 
650 7 |a Peptide Library  |2 NLM 
650 7 |a Platelet Glycoprotein GPIIb-IIIa Complex  |2 NLM 
650 7 |a arginyl-glycyl-aspartic acid  |2 NLM 
650 7 |a 78VO7F77PN  |2 NLM 
700 1 |a Robert, R  |e verfasserin  |4 aut 
700 1 |a Pouns, O  |e verfasserin  |4 aut 
700 1 |a Laroche-Traineau, J  |e verfasserin  |4 aut 
700 1 |a Nurden, A  |e verfasserin  |4 aut 
700 1 |a Peter, K  |e verfasserin  |4 aut 
700 1 |a Little, M  |e verfasserin  |4 aut 
700 1 |a Clofent-Sanchez, G  |e verfasserin  |4 aut 
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