Negative regulator of pluripotent hematopoietic stem cell proliferation in human white blood cells and plasma as analysed by enzyme immunoassay.
This paper describes the analysis, by a highly sensitive and specific enzyme immunoassay (EIA), of AcSDKP, a tetrapeptide recently isolated from fetal calf bone marrow and subsequently purified and identified which substantially inhibits entry into cycle of hematopoietic pluripotent stem cells (CFU-S). This molecule has a marked protective effect in mice during anticancer chemotherapy with phase-specific drugs and plays an essential role in maintaining CFU-S out of cycle in normal mice. Using acetylcholinesterase-AcSDKP conjugate as tracer, rabbit specific antiserum and 96-well microtiter plates coated with a mouse monoclonal anti-rabbit IgG antibody, this EIA allows detection of AcSDKP at 15 fmol levels with a coefficient of variation less than 10% in the 50-500 fmol range. When combined with high-performance liquid chromatography, this assay clearly reveals the presence of this peptide in normal human white blood cells whereas in supernatant from cultured lymphocytes and in plasma the immunoreactive material is distinct from standard AcSDKP.
Pradelles
P
P
Département de Biologie, Commissariat à l'Energie Atomique, CEN/Saclay, Gif sur Yvette, France.
Frobert
Y
Y
Créminon
C
C
Liozon
E
E
Massé
A
A
Frindel
E
E
D000110:Acetylcholinesterase; D000595:Amino Acid Sequence; D000918:Antibody Specificity; D002455:Cell Division / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007124:Immunoenzyme Techniques; D007962:Leukocytes / Q000032:analysis*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000097:blood* / Q000494:pharmacology; D010949:Plasma / Q000032:analysis*
D009842:Oligopeptides; D000110:Acetylcholinesterase; C058504:goralatide