A rapid method for generating and characterizing anti-variable region monoclonal antibodies

Hum Antibodies. 2006;15(4):155-62.

Abstract

The generation of anti-variable region monoclonal antibodies (mAbs) against therapeutic antibodies is essential in the pharmacokinetic/pharmacodynamic (PK/PD) assessments of the drugs in clinical study samples. Sandwich EIA and other methods are typically employed to achieve sensitivity and selectivity for the PK/PD analyses. These assays usually require generation of mAb reagents that bind specifically to the therapeutic mAb candidate in non-competing pair combinations. Thus, large panels of anti-variable region mAbs must be generated in an expeditious manner to increase the probability of success. Previously, we described a novel immunization method using type 1 interferons (IFNs) coupled with an agonistic anti-CD40 mAb to drive immune responses (Staquet et al., Human Antibodies 15 (2006), 61-69). This protocol allows for rapid and robust generation of large panels of anti-variable region mAbs. In order to quickly characterize and efficiently identify optimal anti-variable region antibody pairs early in the hybridoma process using crude supernatants, an inexpensive, high-throughput ELISA method was developed. The ability to rapidly identify appropriate mAb pairs will save resources by eliminating the time-consuming and laborious process of subcloning irrelevant hybridomas.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Antibodies, Monoclonal / administration & dosage
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / pharmacokinetics
  • Antibodies, Monoclonal / therapeutic use
  • B-Lymphocytes
  • Biotechnology / methods
  • CD40 Antigens / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Humans
  • Immunization
  • Immunoglobulin G / administration & dosage
  • Immunoglobulin G / immunology
  • Immunoglobulin Variable Region / immunology*
  • Interferon Type I
  • Mice
  • Mice, Inbred BALB C
  • Time Factors

Substances

  • Antibodies, Monoclonal
  • CD40 Antigens
  • Immunoglobulin G
  • Immunoglobulin Variable Region
  • Interferon Type I