Affiliations: Centocor Research & Development Inc. 145 King of Prussia Rd., Radnor, PA 19087, USA
Correspondence:
[*]
Corresponding author: Centocor Research & Development Inc. 145 King of Prussia Rd., Radnor, PA 19087, USA. Tel.: +1 610 651 6223 direct; Fax: +1 610 651 6798; E-mail: KStaquet@cntus.jnj.com.
Note: [1] Both authors contributed equally to the paper.
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.
Keywords: Antibodies (Ab), Monoclonal antibodies (mAb), anti-variable region monoclonal antibodies, enzyme link immunosorbent assay (ELISA), Pharmacokinetic and pharmacodynamic (PK/PD), B cell proliferative agent, hybridoma, interferons
