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Soluble T cell receptors: detection and quantitative assay in fluid phase via ELISA or immuno-PCR.
- Source :
-
Journal of immunological methods [J Immunol Methods] 1995 Oct 26; Vol. 186 (2), pp. 181-94. - Publication Year :
- 1995
-
Abstract
- 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.
- Subjects :
- Animals
Antibodies, Monoclonal immunology
Antibodies, Monoclonal isolation & purification
Base Sequence
Chromatography, Affinity
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
Cricetinae
Enzyme-Linked Immunosorbent Assay instrumentation
Humans
Mice
Molecular Sequence Data
Osmolar Concentration
Receptors, Antigen, T-Cell, alpha-beta isolation & purification
Receptors, Antigen, T-Cell, gamma-delta analysis
Recombinant Proteins immunology
Sensitivity and Specificity
Solubility
Enzyme-Linked Immunosorbent Assay methods
Polymerase Chain Reaction methods
Receptors, Antigen, T-Cell, alpha-beta analysis
Th2 Cells chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 186
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 7594618
- Full Text :
- https://doi.org/10.1016/0022-1759(95)00129-x