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Variable region differences affect antibody binding to immobilized but not soluble antigen

Variable region differences affect antibody binding to immobilized but not soluble antigen We have examined the antigen binding characteristics of two chimeric IgG 1 antibodies that differ only in the heavy chain variable region. Antibodies 10B and B11 were expressed from two different anti-(Tyr, Glu)-Ala–Lys murine V H genes joined to human IgG 1 constant region genes in a murine anti-(Tyr, Glu)-Ala-Lys heavy chain loss variant hybridoma. The binding characteristics of the antibodies to (Tyr,Glu)-Ala–Lys and to a peptide conjugate, CYYYEEEEY:BSA, were measured in solution and solid phase assays. The antibodies exhibited similar affinities and binding characteristics when assayed in solution assays. However, when we measured binding of antibodies to immobilized antigens, we found that antibody affinity depended on the epitope density in the immobilized immune complexes. The binding of antibody 10B and of B11 to immobilized (Tyr,Glu)-Ala–Lys and to CYYYEEEEY:BSA were similar at high antigen density, but antibody B11 bound less well at lower antigen density. Fab fragments of 10B bound to immobilized (Tyr,Glu)-Ala–Lys and CYYYEEEEY:BSA, but Fab fragments of B11 did not bind to (Tyr,Glu)-Ala–Lys and bound less well to CYYYEEEEY:BSA than 10B Fabs. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Human Antibodies IOS Press

Variable region differences affect antibody binding to immobilized but not soluble antigen

Horgan, Carol; Brown, Kathy; Pincus, Seth H.
Human Antibodies , Volume 3 (3) – Jan 1, 1992
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References (7)

Publisher
IOS Press
Copyright
Copyright © 1992 by IOS Press, Inc
ISSN
1093-2607
eISSN
1875-869X
DOI
10.3233/HAB-1992-3306
Publisher site
See Article on Publisher Site

Abstract

We have examined the antigen binding characteristics of two chimeric IgG 1 antibodies that differ only in the heavy chain variable region. Antibodies 10B and B11 were expressed from two different anti-(Tyr, Glu)-Ala–Lys murine V H genes joined to human IgG 1 constant region genes in a murine anti-(Tyr, Glu)-Ala-Lys heavy chain loss variant hybridoma. The binding characteristics of the antibodies to (Tyr,Glu)-Ala–Lys and to a peptide conjugate, CYYYEEEEY:BSA, were measured in solution and solid phase assays. The antibodies exhibited similar affinities and binding characteristics when assayed in solution assays. However, when we measured binding of antibodies to immobilized antigens, we found that antibody affinity depended on the epitope density in the immobilized immune complexes. The binding of antibody 10B and of B11 to immobilized (Tyr,Glu)-Ala–Lys and to CYYYEEEEY:BSA were similar at high antigen density, but antibody B11 bound less well at lower antigen density. Fab fragments of 10B bound to immobilized (Tyr,Glu)-Ala–Lys and CYYYEEEEY:BSA, but Fab fragments of B11 did not bind to (Tyr,Glu)-Ala–Lys and bound less well to CYYYEEEEY:BSA than 10B Fabs.

Journal

Human Antibodies – IOS Press

Published: Jan 1, 1992

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