J. Sperl et al., SOLUBLE T-CELL RECEPTORS - DETECTION AND QUANTITATIVE ASSAY IN FLUID-PHASE VIA ELISA OR IMMUNO-PCR, Journal of immunological methods, 186(2), 1995, pp. 181-194
To establish the concentration range in which soluble murine T cell re
ceptors (sTCR), derived from the Th2 clone D10, exhibited biological a
ctivity, and to follow production and purification of D10 sTCR, we dev
ised four quantitative immunoassays: three ELISA systems, and an immun
o-PCR assay. The direct ELISA, employed hamster anti-TCR beta monoclon
al antibody (H57), which detects all types of cup alpha beta TCR, rega
rdless 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 cap
ture 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 cup alpha beta TCR, and showed no cross-reactivity when em
ploying two control hamster anti-gamma delta TCR mAbs (GL3 and UC7), o
r with anti-TCR beta and monoclonal hamster IgG as a control antigen.
Further, we demonstrated that in some assays where use of passive bind
ing ELISA plates resulted in a high background, replacement with coval
ent binding ELISA plates resulted in an acceptable low background valu
e. 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 pro
ve valuable in investigating the occurrence and amount of sTCR in vitr
o and in vivo.