The Use of Antibody Affinity Chromatography and Other Methods in the Study of Pregnancy-Associated Proteins
The search for new pregnancy associated proteins has involved many disciplines and included many enzymatic, hormonal, electro-phoretic and immunological techniques (see BOHN, 1976). Enzymatic and hormonal methods are usually specific for proteins of known function or effect, whose nature must, therefore, be defined before the protein can be detected. By contrast, electrophoretic and immunological procedures can be carried out without assumption as to the function of the protein being sought. The broad applicability of electrophoretic and immunological methods is, however, offset by their limited ability to distinguish tissue-specific proteins which exist in low concentration amongst the very wide array of intracellular and extracellular proteins found in virtually all tissues. Thus, immunisation of rabbits with 100-300 mg of protein from extracts of human placenta or human fetal liver yielded precipitating antibodies against the proteins commonly found in adult human serum. After adsorption with solid phase adult human serum protein (AVRAMEAS and TERNYNCK, 1969), only some ten precipitin arcs could be detected by antibody-antigen crossed electrophoresis (AACE). We have used two approaches to reduce the strong antibody response to high concentration serum proteins.
KeywordsAlbumin Urea Carbohydrate Electrophoresis MgCl2
Unable to display preview. Download preview PDF.
- Bohn, H.: Isolation and characterisation of placental specific proteins SP-1 and PP-5. Protides Biol. Fluids 24, 117–124 (1976)Google Scholar
- Bolton, A.E.: Radioiodination techniques. In: Reviews of the radiochemical centre, Vol. XVII, pp. 45–58. Amersham: Radiochemical Centre 1977Google Scholar
- Hobbs, J.R.: Affinity purified antibodies enabling immunoperoxidase reactions. Communicated at the 24th meeting of protides of the biological fluids 1976Google Scholar
- Horne, C.H.W., Bohn, H., Towler, C.M.: Pregnancy-associated α2-glycoprotein. In: Plasma hormone assays in the evaluation of fetal wellbemg. KLOPPER, A. (ed.). Edinburgh: Churchill Livingstone 1976Google Scholar
- Hunter, W.M.: Radioimmunoassay. In: Immunochemistry. WEIR, J. (ed.), Vol. I, pp. 14.1-14.40. Oxford: Blackwell 1976Google Scholar
- Seppälä, M., Ruoslahti, E.: Alpha-fetoprotein. In: Contributions to gynecology and obstetrics. KELLER, P.J. (ed.), pp. 144–186. Basel: Karger 1976Google Scholar
- Sutcliffe, R.G.: The search for new human fetal proteins. Protides Biol. Fluids 24, 543–546 (1976)Google Scholar
- Sutcliffe, R.G., Brock, D.J.H., Nicholson, L.V.B., Dunn, E.: Fetal and uterine-specific antigens in human amniotic fluid. J. Reprod. Fertil. 54 (1978a) (in press)Google Scholar
- Sutcliffe, R.G., Bolton, A.E., Sharp, F., Nicholson, L.V.B., Mackinnon, R.: The purification of human alpha uterine protein (AUP) by antibody affinity chromatography (Submitted) (1978b)Google Scholar