Allotyping for HLA class I using plasma as antigen source.
Immunoadsorption of soluble HLA class I antigens onto immunobeads, one-dimensional iso-electric focusing of these proteins and subsequent immunoblotting allows a biochemical identification of HLA class I allotypes. The distinct protein bands can be clearly attributed to particular HLA antigens and are comparable to those observed after detergent solubilization of membrane-bound HLA antigens. Segregation analysis showed that the biochemically detected pattern of soluble class I gene products followed Mendelian inheritance. However, antigens such as HLA-A1, -A2, -B8, and -B51 were not always clearly detectable, a phenomenon attributable to either different plasma concentrations of these HLA antigens or variable affinity of the monoclonal antibody used to capture class I antigens. These results show that in principle allotyping of HLA class I using plasma as the antigen source is feasible, but with the limitation that some antigens may not be easily detected in some individuals.
Grosse-Wilde
H
H
Institut für Immungenetik, Universitätsklinikum Essen, F.R.G.
Doxiadis
I
I
D015395:Histocompatibility Antigens Class I / Q000302:isolation & purification; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D007525:Isoelectric Focusing; D008280:Magnetics; D010949:Plasma / Q000276:immunology
D015395:Histocompatibility Antigens Class I