Simultaneous HLA Class I and Class II antibodies screening with flow cytometry.
A flow cytometric method of simultaneously screening both HLA Class I and Class II panel reactive antibodies (PRA) was developed using a pool of 30 different Class I and 30 different Class II microbeads coated with purified HLA antigens. The antibodies in the serum that react specifically to the coated HLA antigens are detected by using a FITC-conjugated antibody against human IgG. Percent PRA can be determined by the percentage of microbeads that react positively to the serum. There is no cross-reactivity between the Class I and Class II microbeads. A mixture of Class I and Class II microbeads can be distinguished by their different fluorescent properties on the flow cytometry analysis. Thus, both Class I and Class II PRA can be detected from a single tube reaction.
Pei
R
R
Department of Research, One Lambda, Inc., Canoga Park, CA 91303, USA.
Wang
G
G
Tarsitani
C
C
Rojo
S
S
Chen
T
T
Takemura
S
S
Liu
A
A
Lee
J
J
D000906:Antibodies / Q000097:blood / Q000302:isolation & purification*; D003429:Cross Reactions / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D006801:Humans; D012680:Sensitivity and Specificity
D000906:Antibodies; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II