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Related Experiment Videos

Isotype choice for chimeric antibodies affects binding properties

M M Morelock1, R Rothlein, S M Bright

  • 1Department of Biochemistry, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut 06877-0368.

The Journal of Biological Chemistry
|April 29, 1994
PubMed
Summary
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Murine and chimeric anti-intercellular adhesion molecule 1 (ICAM-1) antibodies bind with equal affinity. However, their avidity differs, impacting how strongly they bind to ICAM-1 on cell surfaces.

Area of Science:

  • Immunology
  • Biochemistry
  • Molecular Biology

Background:

  • Intercellular adhesion molecule 1 (ICAM-1) plays a crucial role in cellular interactions.
  • Murine antibody BIRR1 recognizes ICAM-1.
  • Chimeric antibodies combine murine and human regions to potentially improve therapeutic properties.

Purpose of the Study:

  • To investigate the binding characteristics of chimeric antibodies derived from BIRR1.
  • To compare the binding affinity and avidity of chimeric antibodies to ICAM-1 with the original murine antibody.

Main Methods:

  • Construction of chimeric antibodies with human constant regions and murine variable regions.
  • Solid-phase competitive enzyme-linked immunosorbent assay (ELISA) to assess binding competition.
  • Papain digestion to generate Fab fragments.

Related Experiment Videos

  • Size exclusion high-performance liquid chromatography (HPLC) for solution-phase binding assays.

    • Main Results:

    • Chimeric antibodies exhibited differential binding abilities to ICAM-1.
    • Binding competition order: BIRR1 = cIgG1 > cIgG4 > cIgG2.
    • Fab fragments of murine and chimeric antibodies showed equivalent binding constants.
    • Intact antibodies demonstrated equivalent binding constants for their Fab arms but differing avidities.

    Conclusions:

    • Murine and chimeric anti-ICAM-1 antibodies possess equivalent binding affinities.
    • Differences in binding competition are attributed to variations in avidity, not affinity.
    • Avidity differences influence the overall strength of antibody binding to cellular ICAM-1.