High-performance liquid affinity chromatography of liver plasma membrane proteins

Josić, Djuro; Hofmann, Werner; Habermann, R.; Becker, Andreas; Reutter, Werner

doi:10.1016/s0021-9673(01)84988-8

Cited by 19 publications

(3 citation statements)

References 11 publications

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“…Monoclonal antibodies (hMbs) have also been used to identify other molecules that are specific for a single domain (4)(5)(6). In particular, several molecules of unknown function have been recently described on the bile canalicular cell surface (7)(8)(9)(10)(11)(12).…”

mentioning

confidence: 99%

Identification of the bile canalicular cell surface molecule GP110 as the ectopeptidase dipeptidyl peptidase IV: An analysis by tissue distribution, purification andN-terminal amino acid sequence

et al. 1990

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This paper describes the tissue distribution, purification and N-terminal amino acid sequence of the bile canalicular cell surface molecule dipeptidyl peptidase IV. Immunoperoxidase staining of cryostat sections of rat liver with a monoclonal antibody, Medical Research Council OX-61, indicated specific binding to hepatocyte bile canalicular domains and brush borders of bile ducts. Additional staining was seen in other epithelial brush borders (small intestine, kidney, colon, pancreatic duct); acinar structures in salivary glands; endothelial structures and T cell areas in thymus, spleen and lymph node. The tissue distribution suggested that monoclonal antibody OX-61 binds to the ectoenzyme dipeptidyl peptidase IV. This was confirmed by depletion of dipeptidyl peptidase IV activity from tissue homogenates by monoclonal antibody OX-61 coupled to Sepharose. The molecule recognized by OX-61 was then purified from liver and kidney by monoclonal antibody affinity chromatography. The molecule had a molecular weight of 110 kD under reducing conditions. The purified molecule was subsequently analyzed for amino acid composition and N-terminal amino acid sequence. Thirty-one N-terminal amino acids were sequenced and indicated identity with part of the predicted N-terminus of the previously cloned bile canalicular molecule GP110. On review, other similarities between dipeptidyl peptidase IV and GP110 were detected: molecular weight, deglycosylated form and metabolic half-life. Finally, the recent cloning of dipeptidyl peptidase IV permitted a comparison between the molecule recognized by monoclonal antibody OX-61, GP110 and dipeptidyl peptidase IV. It is concluded that these three molecules are almost certainly identical.

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confidence: 99%

Identification of the bile canalicular cell surface molecule GP110 as the ectopeptidase dipeptidyl peptidase IV: An analysis by tissue distribution, purification andN-terminal amino acid sequence

et al. 1990

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“…It follows that a 20 χ 4.6 mm column, containing about 200 mg of silica, has bound approximately 11 mg of antibody. Such "in situ" immobilizations of different ligands have been described before (2,22,23). The first commercially available columns, which are packed with an epoxy-activated support, are less effective because of the rather low reactivity of the epoxy groups.…”

Section: Immunoaffinity Hplcmentioning

confidence: 97%

Isolation of Immunoglobulins and Their Use in Immunoaffmity HPLC

Josić¹,

Hofmann²,

Habermann³

et al. 1988

Clinical Chemistry and Laboratory Medicine

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Summary:For the isolation of monoclonal and polyclonal antibodies different high performance liquid chromatography (HPLC) and high performance affinity chromatography (HPAC) methods were investigated. Specially designed "mixed-bed" ion-exchange and hydroxylapatite columns as well as hydrophobic interaction columns were efficiently applied to the isolation of monoclonal antibodies. When these methods are used for the isolation of polyclonal antibodies from antiserum, the sample has to be pre-treated, e. g. by removal of serum albumin.Protein A HPAC is an easy method and quick to handle, especially for the preparative isolation of antibodies. The antibodies that do not bind to protein A, can be purified by protein G HPAC. If this method cannot be used because of the rather extreme elution conditions, hydroxylapatite, ion-exchange or hydrophobic interaction HPLC have to be considered as alternatives.We further concentrate on immunoaffinity HPLC with immobilized antibodies. This method has proved to be very effective for one-^step isolation of antigens, even from very complex samples such as plasma membrane extracts. The problem with immunoaffinity HPLC is the quick deterioration of the columns, caused by increasing denaturing of the immobilized antibodies during elution. In order to solve this problem, an indirect method is recommended for analytical immunoaffinity HPLC. For this purpose, the antibodies are bound to a protein A HPAC column. The solution containing the antigens is then applied. After washing, the antigenantibody complex is eluted from the column.

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“…Isolation of ral serum transferrin by immunoaffinity high performance liquid chromatography Serum was obtained by coagulation of blood at room temperature and subsequent centrifugation at 1000g for 15 min. Albumin and IgG were removed from the serum by chromatography on a afifi-blue-gel column (BioRad, München, F. R. G.) or on a Eupergit-protein A (30N) 4.6 60 mm HPLC coluinn, äs previously described (10). Subsequently, transferrin was isolated by immunoaffinity high performance liquid chromatography using polyclonal rabbit anti-transferrin antibodies (Cappel, CochranviUe, PA. 19330, U. S.…”

Section: Polyacrylamide Gel Electrophoresismentioning

confidence: 99%

Half-Lives of L-[35S]Methionine and L-[3H]Fucose of Transferrin in the Serum of Rats

Josić

Tauber²,

Hofmann³

et al. 1987

Clinical Chemistry and Laboratory Medicine

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The half-lives of 35 S-labelled L-methionine and of 3 H-labelled L-fucose of serum transferrin of rats were measured in pulse-chase experiments in vivo. Both L-[ 35 S]methionine and L-[6-3 H[fucose disappeared from transferrin with nearly the same half-lives of 33.8 h and 36.5 h, respectively. The data show that in this major serum glycoprotein peripheral carbohydrates and the protein moiety are degraded äs a unit.

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High-performance liquid affinity chromatography of liver plasma membrane proteins

Cited by 19 publications

References 11 publications

Identification of the bile canalicular cell surface molecule GP110 as the ectopeptidase dipeptidyl peptidase IV: An analysis by tissue distribution, purification andN-terminal amino acid sequence

Identification of the bile canalicular cell surface molecule GP110 as the ectopeptidase dipeptidyl peptidase IV: An analysis by tissue distribution, purification andN-terminal amino acid sequence

Isolation of Immunoglobulins and Their Use in Immunoaffmity HPLC

Half-Lives of L-[35S]Methionine and L-[3H]Fucose of Transferrin in the Serum of Rats

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