AN ENZYME-LINKED-IMMUNOSORBENT-ASSAY AND REFERENCE RANGES FOR BISPHOSPHOGLYCERATE MUTASE IN HUMAN ERYTHROCYTES

We established an enzyme-linked immunosorbent assay (ELISA) system for the determination of human bisphosphoglycerate mutase (BPGM) protein content in human erythrocytes using a polyclonal anti-BPGM antibody, w e determined reference ranges for BPGM protein content, synthase activ ity, and specific activity in human erythrocytes. We produced a recomb inant human BPGM (rBPGM) by gene manipulation using E. coli and then o btained the polyclonal antibody by immunizing rabbits with purified rB PGM. The reproducibility of the ELISA was in an acceptable range with a coefficient of variation under 1.5%. The ELISA was reliable in the r ange of 0.1 to 10 ng/mL. The polyclonal anti-rBPGM antibody did not sh ow any cross-reaction with recombinant human B type phosphoglycerate m utase, which is highly homologous to rBPGM. The ELISA was found to be practical for the determination of BPGM protein content in human eryth rocytes. The mean BPGM protein content was 56.3 +/- 9.7 mu g/mL in who le blood (mean +/- SD, n = 50). The ELISA can be used to examine vario us hematologic disorders with abnormal red cell size and cell counts, and to detect BPGM enzymopathy in human erythrocytes. J. Clin. Lab. An al. 12:263-267, 1998. (C) 1998 Wiley-Liss, Inc.