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M9620514.TXT
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1996-02-26
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Document 0514
DOCN M9620514
TI Soluble T cell receptors: detection and quantitative assay in fluid
phase via ELISA or immuno-PCR.
DT 9602
AU Sperl J; Paliwal V; Ramabhadran R; Nowak B; Askenase PW; Department of
Medicine, Yale University School of Medicine, New; Haven, CT 06520-8013,
USA.
SO J Immunol Methods. 1995 Oct 26;186(2):181-94. Unique Identifier :
AIDSLINE MED/96062061
AB To establish the concentration range in which soluble murine T cell
receptors (sTCR), derived from the Th2 clone D10, exhibited biological
activity, and to follow production and purification of D10 sTCR, we
devised four quantitative immunoassays: three ELISA systems, and an
immuno-PCR assay. The direct ELISA, employed hamster anti-TCR beta
monoclonal antibody (H57), which detects all types of alpha beta TCR,
regardless of their variable regions, and had a detection limit of about
6 ng/ml sTCR. The indirect sandwich ELISA employed anti-V beta 8 as
capture antibody, and had a detection limit of 600 pg/ml. With the
direct sandwich ELISA, that also employed anti-V beta 8, TCR
concentrations as low as 100 pg/ml could be detected. The ELISA assays
were specific for soluble alpha beta TCR, and showed no cross-reactivity
when employing two control hamster anti-gamma delta TCR mAbs (GL3 and
UC7), or with anti-TCR beta and monoclonal hamster IgG as a control
antigen. Further, we demonstrated that in some assays where use of
passive binding ELISA plates resulted in a high background, replacement
with covalent binding ELISA plates resulted in an acceptable low
background value. With the immuno-PCR assay, concentrations of sTCR as
little as 0.8 pg/ml could be detected. In summary, the assays described
here may prove valuable in investigating the occurrence and amount of
sTCR in vitro and in vivo.
DE Animal Antibodies, Monoclonal/IMMUNOLOGY/ISOLATION & PURIF Base
Sequence Chromatography, Affinity Chromatography, High Pressure Liquid
Chromatography, Ion Exchange Comparative Study Enzyme-Linked
Immunosorbent Assay/INSTRUMENTATION/*METHODS Hamsters Human Mice
Molecular Sequence Data Osmolar Concentration Polymerase Chain
Reaction/*METHODS Receptors, Antigen, T-Cell,
alpha-beta/*ANALYSIS/ISOLATION & PURIF Receptors, Antigen, T-Cell,
gamma-delta/ANALYSIS Recombinant Proteins/IMMUNOLOGY Sensitivity and
Specificity Solubility Support, U.S. Gov't, P.H.S. Th2
Cells/*CHEMISTRY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).