Determination of lectin-sugar dissociation constants by agarose affinity electrophoresis.

Agarose crossed affinity electrophoresis (aff-EP) was employed for the determination of lectin-sugar dissociation constants (Ki). In the first dimension of the aff-EP increasing amounts of sugar (alpha-methyl-D-mannoside) were added to a given concentration of lectin (concanavalin A). Then the electrophoresis was run with alpha 1-acid glycoprotein, alpha 1-antitrypsin and alpha-fetoprotein as markers of lectin-sugar interactions.

Studies on microheterogeneity of acute-phase proteins in rheumatoid arthritis by using crossed affinoimmuno-electrophoresis with free concanavalin A.

Microheterogeneity of two acute-phase proteins: orosomucoid (alpha1-acid glycoprotein, AGP) and alpha 1-antichymotrypsin (ACHT) were studied in the sera of 48 patients with rheumatoid arthritis and 12 healthy individuals. Each rheumatoid patient was assigned to one of four activity grades. Cross affinoimmunoelectrophoresis (aff-EP) with free Concanavalin A (Con A) revealed three microheterogeneity variants of AGP and four microheterogeneity forms of ACHT.

The effect of C-reactive protein on the PHA-induced proliferation of human peripheral blood lymphocytes.

The effect of CRP purified from ascites fluid originated from cancer patients on human peripheral blood lymphocytes was investigated. CRP was purified using 4-step procedure including absorption on Sepharose 4B, phosphocholine-Sepharose, DE-52 ion exchange chromatography and gel filtration on Sephacryl S-200. CRP was able either to inhibit or to enhance PHA-induced lymphocyte proliferation in vitro.

Comparison of three immunoassays for C-reactive protein determination.

Three widely used immunoassays for C-reactive protein (CRP) determination (RID. RIEP and LN--Behring) were compared. According to the CRP calcium dependent structural changes or ligand binding properties two buffer systems, Ca2+ rich and Ca2+ depleted, were used in all methods.

Lectin inhibition system for determination of concanavalin A glycoprotein complexes dissociation constants in agarose affinity electrophoresis.

A new system for lectin-glycoprotein complexes dissociation constants (K) determination is presented. The system is based on agarose affinity electrophoresis where equal lectin (Con A) concentrations are inhibited by variable specific sugar (alpha-methyl-mannoside) amounts. Moreover, the system allows lectin-sugar inhibition constants (Ki) studies.

Microheterogeneity forms of alpha-fetoprotein present in amniotic fluid.

The microheterogeneity forms of human alpha-fetoprotein present in amniotic fluid were studied in affinity immunoelectrophoresis using two lectins: concanavalin A and Lens culinaris agglutinin. Ninety-seven samples of amniotic fluid collected throughout gestation were obtained from pregnancies in which there was no fetal malformation. Two samples of amniotic fluid from pregnancies complicated by anencephaly and one sample obtained from an empty gestational sac were also studied.

Microheterogeneity of alpha-fetoprotein in patient serum as demonstrated by lectin affino-electrophoresis.

Microheterogeneity of alpha-fetoprotein (AFP) present in the sera of 76 patients was studied by lectin affino-immunoelectrophoresis. Seventeen patients had benign liver disorders and the remaining 59 patients were treated for primary or secondary liver cancer or yolk sac tumour. By means of Con A, AFP was divided into two variants, while lentil lectin (LCA) made it possible to separate AFP in three variants.

The in vitro production of alpha--fetoprotein variants by human fetal organs.

We have demonstrated the immunological identity of alpha-fetoprotein (AFP) produced in vitro by various fetal organs such as the yolk sac, liver, intestine and kidney. AFP produced in vitro by these organs, present in fetal serum and in samples of amniotic fluid was, however, heterogeneous when studied by lectin affinity immunoelectrophoresis. Results obtained with Concanavalin A were comparable to those obtained with Lentil agglutinin, but Concanavalin A separated AFP into two variants only, while Lentil agglutinin recognised three AFP variants.