| Hemangiopoietin is a novel protein first detected in the urine of aplastic anemia patients. Its gene has been cloned and proved to be a new splicing variant of proteoglycan 4. Its recombinant form has been found to have the ability to stimulate the proliferation of hematopoietic and endothelial stem/progenitor cells significantly in vitro in both human and murine systems. When rh-HAPO was administered to normal or irradiated mice, the result was either a significant increase in bone marrow hematopoietic cells or an enhanced survival rate and accelerated hematopoiesis. To aid in the basic and clinical research of HAPO, we made both monoclonal and polyclonal antibodies against HAPO and constructed an ELISA kit with them to detect HAPO level in the blood serum, urine or culture supernatant. We also investigated the existence of natural HAPO in the human blood serum and human fetal liver by SDS-PAGE and western blot with our monoclonal antibody. Among the MoAbs we screened out, three were IgG1 and two were IgM. The subtype of their light trains were all belonged to K.The relative affinities of the three IgGl form MoAbs detected with indirect ELISA were 3.06× 109 M-1, 6.07×108 M-1 and 1.71 × 1010 M-1 respectively and they can all be used in both frozen and paraffin embedded tissue sections. The epitope recognition of the three IgGl MoAb was conformational independent confirmed by SDS-PAGE with partly degradated HAPO... |
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