Isolierung von Komponenten menschlicher Magenschleimhaut-Pepsinogene und -Cathepsine mit präparativer Polyacrylamidgelelektrophorese zur Herstellung spezifischer Immunseren

Abstract

Zusammenfassung: Mit der präparativen Polyacrylamidgelelektrophorese wurden aus menschlichen Magenschleimhautextrakten Komponenten der immunchemisch verschiedenen Pepsinogengruppen I, II und des Magenschleimhautcathepsins gewonnen. Die Immunisierung von Schafen und Kaninchen ergab spezifische Immunseren, die gegen die drei verschiedenen Systeme der Magenschleimhautproteasen gerichtet waren. Isolation of components ofpepsinogensl, II and mucosal cathepsin of lutman gastric mucosa by preparative poly· acrylamide gel electrophoresis, and the preparation of specific antiseraSummary: Components of pepsinogens I, II and mucosal cathepsin from human gastric mucosa were purified from gastric extracts by preparative polyacrylamide gel electrophoresis. Specific antisera directed against three different systems of gastric proteases were raised by immunization of sheep and rabbits. Einführung

Charakterisierung von menschlichem Pepsin I gewonnen aus gereinigtem Magenpepsinogen I

“…According to previous studies [36,16,28] the subsets within one pepsinogen group are immunochemically identical.…”

“…Specific antisera (against PgI and PgII) were used in the double gel diffusion technique of Ouchterlony, in the quantitative radial diffusion technique of Mancini and in immunoelectrophoresis (IE) as described earlier [28]. The agarose enzyme electrophoresis (AEE) was described elsewhere [28].…”

“…The agarose enzyme electrophoresis (AEE) was described elsewhere [28]. The polyacrylamid gel electrophoresis (PAGE) was performed according to the Allen-procedure [1] using a discontinuous voltage gradient and discontinuous gel concentration with an Ortec gel electrophoresis system [28].…”

“…Only by the recent application of immunoadsorption techniques using specific immunsera [28] could PgI and PgII be purified from human gastric mucosa fulfilling biochemical and immunochemical criteria of purity [66,23].…”

Radioimmunological Quantitation of Human Group-II Pepsinogens

“…Purified group I pepsinogens showed no response up to concentrations o f 100 ug/ml. In apparently healthy donors, we observed mean plasma concentrations o f 20.3 ng/ml (males) and o f 15.5 ng/ml (females).Gastric acid protease zymogens have been identified in human serum and it has been claimed that their concentration may reflect the secretory status o f the gastric mucosa (4).Immunochemical analysis reveals a striking heterogeneity o f the gastric acid protease system, this being composed o f two immunologically different groups (6, 10) called pepsinogen I (Pgl) and pepsinogen II (Pgll) (13), which by them selves can be resolved into 4 -5 components differing in electrophoretic mobility (9,11,12,21). Group I and group II pepsinogens have been localized by immunochemical methods in the body o f the stomach; Pgll has also been found in the antrum and the upper part o f the duodenum (14, 15).…”