Isolierung des schwangerschaftsspezifischen ?1-Glykoproteins (SP1) und antigenverwandter Proteine durch Immunadsorption

Abstract

Purification of human pregnancy-specific beta1-glycoprotein (SP1) and antigenically related proteins of sub-human primates (chimpanzee, rhesus monkey, cynomolgus and baboon) was achieved by means of an immunoadsorbent technique. The immunoglobulins of a rabbit antiserum to human SP1 were isolated on DEAE-cellulose and coupled to CNBr-activated Sepharose. This immunoadsorbent was used to bind human SP1, respectively monkey proteins immunochemically related to SP1 from placental extract fractions. After extensive washing the proteins were eluted by an acidic glycine buffer. Contaminating serum proteins could be removed by chromatography on hydroxyapatite columns. With this method it was possible to obtain SP1 and the antigenically related proteins of monkeys in good yield and in highly purified form. The proteins thus isolated from human and sub-human primate placentae were compared in their physicochemical and immunochemical properties. The amino acid and carbohydrate compositions of human SP1 and rhesus SP1 have been determined. In a biological test certain inhibitory effect of human SP1 on the mixed leukozyte culture (MLC) could be demonstrated.

Immunological Relationships of Human and Subhuman Primate Pregnancy-Associated Plasma Proteins

“…They also found complex gel diffusion pat terns with various primate preparations. This was even more complex in the case of rhesus monkey placental PAPP-C reacting with an tiserum prepared against purified rhesus PAPP-C [32], It was clear that PAPP-B was relatively difficult to visualize by these various gel dif fusion technics, and it is probable that this is due to the relatively low concentration of this protein in pregnancy serum. Because of this, more sensitive methods of assay will be required to analyze its cross-reactivities with pregnancy primate analogues more precisely.…”

Radioimmunoassay of SP1(Pregnancy-specificβ1-glycoprotein) in maternal blood and in amniotic fluid in normal and pathologic pregnancies

“…purified from placenta [4] in 10 μΐ of phosphate buffered saline were mixed with 0.5 m Ci lodide in 10 μΐ of a 125-J-Na-solution (Behringwerke AG, Marburg, FRG; 8-15 Ci 125 J per mg J) and with a Suspension of l Mg Lactoperoxidase (B.C. 1.11.1.7, BOEHRINGER, Mannheim, FRG) in 10 μΐ PBS.…”

Trophoblast-Specific Beta₁-Glycoprotein as a Marker for Pregnancy and Malignancies

“…More recently, Bohn et al (19) used an immunoabsorbent technique for the preparation of the purity TSG from human and sub-human primates placenta extracts. The immunoglobulins against TSG were isolated on DEAE-cellulose and coupled to CHBr-activated Sepharose.…”