Method for detection of anti-HLA antibodies

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 71, 435 794, 435 23, 435 4052, 436507, 436175, 436176, 436811, 436825, G01N 33533

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active

059622393

ABSTRACT:
An improved method for detecting even low titer HLA antibodies in proposed organ transplant recipients employs flow cytometry crossmatching (FCXM) on pronase-treated B-cells and T-cells of the donor. Two-color FCXM is preferably employed. The peripheral blood lymphocytes, after pronase digestion, are maintained under conditions to suppress Fcy R receptor regeneration, combined with sera of the proposed transplant recipient, and tested against control sera, using fluorescent reporting complexing agents. Use of pronase digested lymphocytes permits the assay to distinguish between normal or irrelevant IgG B cell binding and immunologically important HLA antibody binding.

REFERENCES:
Lobo et al., Transplant International, vol. 8, No. 6, pp. 472-480, 1995.

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