The microheterogeneity of serum α1‐antichymotrypsin revealed by interaction with concanavalin A in crossed immunoaffinoelectrophoresis and in affinity chromatography

In crossed immunoaffinoelectrophoresis with free concanavalin A in the first dimension, human serum α1‐antichymotrypsin, purified or in whole serum, exhibited four peaks in presence of 0.02 M α‐methyl‐D‐glucoside added to the second‐dimensional gel. α1‐Antichymotrypsin purified from the serum of a single healthy donor was separated by affinity chromatography into three fractions on a laboratory‐prepared concanavalin A‐Sepharose 4B column: a pass‐through fraction, a retarded fraction and a bound fraction, eluted from the column on addition of sugar to the buffer. These three fractions were analyzed by crossed immunoaffinoelectrophoresis and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and isoelectric focusing before and after desialylation. The results of electrophoresis as well as chemical analyses indicate that these microheterogeneous forms carry glycans with decreasing degrees of branching from the concanavalin A‐pass‐through form to the concanavalin A‐bound form. This approach represents a first step towards the elucidation of the molecular basis of the microheterogeneity of α1‐antichymotrypsin.

[1]  E. Hachulla,et al.  Alpha 1-antichymotrypsin microheterogeneity in crossed immunoaffinoelectrophoresis with free concanavalin A: a useful diagnostic tool in inflammatory syndrome. , 1988, Clinical chemistry.

[2]  M. Miquel,et al.  Effects of severe burns on glycan microheterogeneity of four acute phase proteins. , 1987, Clinica chimica acta; international journal of clinical chemistry.

[3]  G. Lamblin,et al.  Action of trifluoromethanesulfonic acid on highly glycosylated regions of human bronchial mucins. , 1986, Carbohydrate research.

[4]  M. Rabaud,et al.  Human serum α1‐antichymotrypsin is an inhibitor of pancreatic elastases , 1985 .

[5]  J. Hansen,et al.  The microheterogeneity components of orosomucoid and the dissociation constants and mobilities of concanavalin A/corosomucoid complexes in crossed affinoimmunoelectrophoresis with free concanavalin A , 1984 .

[6]  K. Robson,et al.  Sequence homology between human alpha 1-antichymotrypsin, alpha 1-antitrypsin, and antithrombin III. , 1983, Biochemistry.

[7]  I. Kushner.  THE PHENOMENON OF THE ACUTE PHASE RESPONSE * , 1982, Annals of the New York Academy of Sciences.

[8]  R. Hawkes,et al.  A dot-immunobinding assay for monoclonal and other antibodies. , 1982, Analytical biochemistry.

[9]  T. Shimamura,et al.  Enhancement by α-1-antichymotrypsin of antibody response in vivo , 1981 .

[10]  A. Laine,et al.  Purification and characterization of from human pleural fluid and human serum , 1981 .

[11]  P. Arnaud,et al.  Isolectric patterns of human alpha1‐ antichymotrypsin (A1AChy) and A1AChy‐protease complexes , 1981 .

[12]  Jean‐Pierre Martin,et al.  True and false glycoprotein microheterogeneity observed with lectin crossed affinoimmunoelectrophoresis of inter‐alpha‐trypsin‐inhibitor , 1980 .

[13]  J. Kerckaert,et al.  Affinity differences between some commercially available immobilized Con A with rat alpha-fetoprotein. , 1980, Biochemical and biophysical research communications.

[14]  E. Berger,et al.  The microheterogeneity of human plasma alpha1-acid glycoprotein. , 1980, Hoppe-Seyler's Zeitschrift fur physiologische Chemie.

[15]  J. Travis,et al.  Human alpha-1-antichymotrypsin: interaction with chymotrypsin-like proteinases. , 1978, Biochemistry.

[16]  J. Kerckaert,et al.  Characterization and isolation of nine rat alpha-fetoprotein variants by gel electrophoresis and lectin affinity chromatography. , 1977, Biochemical and biophysical research communications.

[17]  O. Bjerrum,et al.  Detection of Biospecific Interaction during the First Dimension Electrophoresis in Crossed Immunoelectrophoresis * , 1975 .

[18]  P. Cuatrecasas,et al.  A simplified method for cyanogen bromide activation of agarose for affinity chromatography. , 1974, Analytical biochemistry.

[19]  T. Bøg‐Hansen.  Crossed immuno-affinoelectrophoresis. An analytical method to predict the result of affinity chromatography. , 1973, Analytical biochemistry.

[20]  W. C. Breckenridge,et al.  Analysis of monosaccharides by gas-liquid chromatography of the O-methyl glycosides as trifluoroacetate derivatives. Application to glycoproteins and glycolipids. , 1972, Journal of chromatography.

[21]  C. Laurell,et al.  Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. , 1966, Analytical biochemistry.