Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P00790 (PGA)
 2,475 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Four different proteases were screened for their capability of selectively digesting murine monoclonal IgGl to obtain active F(ab)2. For the screening, a series of five different mouse monoclonal antibodies (IgGl, k) was used, recognizing different tumor-associated antigens and currently used for radioimmunoimaging studies. The enzymes (pepsin, bromelain, ficin and elastase) showed different fragmentation capability and the fragments obtained showed different stability and immunoreactivity. No digestion was noticed using elastase. Pepsin gave discontinuous results, in that its activity ranged from reduction of IgG to small inactive fragments to an inability to digest the immunoglobulin. Pepsin activity was strongly pH-dependent and immunoreactivity of the obtained fragments was not always conserved. Bromelain and, in particular, ficin gave excellent results. Digestion was always rapid and stable, all five MAbs were reduced to F(ab)2 in a comparable time range and with high yields. Moreover, ficin-obtained F(ab)2 showed a highly conserved immunoreactivity. Therefore, ficin was selected as the murine monoclonal IgGl digestion enzyme to obtain active bivalent antibody fragments. The digestion procedure gave a uniform result for all five different MAbs and was easily scaled up to produce hundreds of milligrams of F(ab)2.
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PMID:A new enzymatic method to obtain high-yield F(ab)2 suitable for clinical use from mouse IgGl. 201 Nov 30

In order to maximize staining, modifications of immunostaining methods have included proteolytic enzyme digestion of tissue. The authors performed a study of the effect of ficin in 110 paraffinized specimens, including tonsil, lymph nodes, benign vascular and nerve sheath tumors, and various carcinomas and sarcomas. This agent was compared with pepsin and bromelain, as alternative proteases. A panel of monoclonal and polyclonal antibodies was used, with and without previous digestion by ficin, pepsin, and bromelain. A score was assigned to each stain, based on the number and intensity of reactive cells. Ficin enhanced staining markedly in immunostains with antibodies to keratin and Factor VIII-related antigen (F8RAG). Conversely, it abolished staining for LN-2 (a lymphoid marker) and weakened reactivity for S-100 in nerve sheath tumors. Bromelain produced similar results, except that it enhanced S-100. Pepsin was comparatively less active than ficin and bromelain overall but did produce the greatest amplification of vimentin staining in sarcomas. Digestion with any of the three enzymes failed to influence reactivities of leukocyte common antigen, UCHL-1 (a lymphoid marker), alpha-1-antichymotrypsin, carcinoembryonic antigen, epithelial membrane antigen, and blood group isoantigens. These results may reflect a dissimilar recognition of peptide targets in some antigenic proteins, by ficin, bromelain, and pepsin. Hence, one enzymatic agent is unlikely to produce optimal staining for all determinants. With this proviso, however, ficin appeared to be the best general enhancer for antigens known to require vigorous digestion (e.g., keratin; F8RAG) for optimal reactivity in paraffin sections.
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PMID:The use of proteolysis with ficin, for immunostaining of paraffin sections. A study of lymphoid, mesenchymal, and epithelial determinants in human tissues. 245 44

The effects of a number of crystalline and highly purified enzymes on elementary bodies of vaccinia are reported. These effects have been followed by determination of amino nitrogen, staining reaction, and studies of infectivity. Pepsin, at a pH which inactivates the virus, results in its solution and rapid release of amino nitrogen. Crystalline trypsin, chymotrypsin, carboxypeptidase, and ribonuclease are without appreciable effect on the virus. Papain within a short time produces profound alteration in the staining reaction of the elementary body with release of amino nitrogen accompanied by complete inactivation of the virus. This reaction is not shared by crystalline ficin, another plant papain, or by cathepsin, an intracellular proteinase analogous to plant papains but of animal origin.
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PMID:CONSTITUENTS OF ELEMENTARY BODIES OF VACCINIA : III. THE EFFECT OF PURIFIED ENZYMES ON ELEMENTARY BODIES OF VACCINIA. 1987 Oct 53