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Clinical Chemistry 46: 137-139, 2000;
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(Clinical Chemistry. 2000;46:137-139.)
© 2000 American Association for Clinical Chemistry, Inc.

Letters

Abbreviated Direct and Indirect ELISAs: A New and Simple Format

John G. Lewisa,1

a Author for correspondence. Fax 64-3-3640-889; e-mail johnL2@chhlth.govt.nz.

Peter A. Elder1

1 Steroid and Immunobiochemistry Laboratory, Canterbury Health Laboratories, Clinical Biochemistry Unit, P.O. Box 151, Christchurch, New Zealand


To the Editor:

Many laboratories, including our own, perform ELISAs using immobilized antibody or antigen. A common format is to coat the ELISA plate with antibody for direct assays, or antigen for indirect assays. ELISA plates are subsequently "blocked" to prevent any further nonspecific absorption.

Direct assays often are performed by incubation of antibody-coated microtiter plate wells with calibrators or patient samples and then washing the plate and detecting bound antigen with a mouse monoclonal antibody, followed by washing and the addition of anti-mouse immunoglobulin-peroxidase, further washing, and the addition of substrate.

We compared our direct sandwich ELISA for sex hormone-binding globulin (SHBG), using this format (1), to a procedure that differs in that the supernatant contains both monoclonal antibody to SHBG and the anti-mouse immunoglobulin-peroxidase (1:20 and 1:500, respectively; cat. no. NA 931; Amersham). The combined antibodies are then added to washed polyclonal anti-SHBG antibody-coated ELISA plate . . . [Full Text of this Article]


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