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).