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Bioanalysis. 2012 Feb;4(3):241-8. doi: 10.4155/bio.11.312.

A novel, high-sensitivity and drug-tolerant sandwich immunoassay for the quantitative measurement of circulating proteins.

Author information

1
Eli Lilly & Company, Indianapolis, IN 46285, USA. sloan_john_harrison@lilly.com

Abstract

BACKGROUND:

Accurate measurement of a total protein target (free plus bound) is essential to optimize dose selection for monoclonal antibody drugs. Herein, we describe a novel sandwich immunoassay format in which the biotherapeutic antibody itself serves as the primary detection antibody. A signal is then generated through the addition of a labeled secondary antibody that recognizes the biotherapeutic antibody. The secondary antibody is conjugated with ruthenium to facilitate electrochemiluminescent analysis.

RESULTS:

Data are presented from the analysis of two protein biomarkers having disparate size and structure; a 4.5 kDa peptide and a 60 kDa protein. In both cases, validated, highly specific assays were developed and shown to be tolerant to elevated levels of the therapeutic monoclonal antibody in question.

CONCLUSION:

Our novel format allows drug-tolerant measurement of soluble protein biomarkers targeted by monoclonal antibodies when only two independent epitopes for antibody binding are available and one is recognized by the therapeutic antibody.

PMID:
22303828
DOI:
10.4155/bio.11.312
[Indexed for MEDLINE]
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