Specific and sensitive detection of purified HLA molecules in an ELISA using mouse, rabbit and human anti-HLA antibodies.

An ELISA detecting anti-HLA antibodies of rabbit, mouse or human origin was developed using plates coated with HLA molecules purified on affinity columns. The sensitivity of the assay was optimal when coating was performed in PBS, pH 7.8 at 4 degrees C for 6-16 h and using a serum incubation period of 16 h at 4 degrees C. The optimum protein concentration for coating was estimated to be 1 micrograms/ml. With monoclonal anti-HLA sera, antipeptide antibodies from mice or rabbit and human alloantisera, this method appeared to be highly sensitive, very specific and reproducible.