Purification of F(ab′) 2 anti-snake venom by caprylic acid: A fast method for obtaining IgG fragments with high neutralization activity, purity and yield
Pooled horse plasma containing antibodies against Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab′) 2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab′) 2B purified by precipitation...
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| Veröffentlicht in: | Toxicon (Oxford) 1989, Vol.27 (3), p.297-303 |
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| container_title | Toxicon (Oxford) |
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| creator | dos Santos, M.C. D'Império Lima, M.R. Furtado, G.C. Colletto, G.M.D.D. Kipnis, T.L. Dias da Silva, W. |
| description | Pooled horse plasma containing antibodies against
Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab′)
2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab′)
2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab′ fragments were obtained by reduction and alkylation of F(ab′)
2B. The anti-whole
C. d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab′)
2B], 4800 [F(ab′)
2M] and 3200 (Fab′B). Immunochemical analysis of these fragments by SDS gel electrophoresis, Western blot and by double immunodiffusion revealed that the solution containing. F(ab′)
2M was free of IgG and of other plasma proteins, whereas that containing F(ab′)
2B was not. One milligram of either F(ab′)
2B, F(ab′)
2M or Fab′B was able to neutralize respectively 20.7 μg, 20.2 μg and 13.8 μg of
C. d. terrificus venom. |
| doi_str_mv | 10.1016/0041-0101(89)90177-3 |
| format | Article |
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Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab′)
2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab′)
2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab′ fragments were obtained by reduction and alkylation of F(ab′)
2B. The anti-whole
C. d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab′)
2B], 4800 [F(ab′)
2M] and 3200 (Fab′B). Immunochemical analysis of these fragments by SDS gel electrophoresis, Western blot and by double immunodiffusion revealed that the solution containing. F(ab′)
2M was free of IgG and of other plasma proteins, whereas that containing F(ab′)
2B was not. One milligram of either F(ab′)
2B, F(ab′)
2M or Fab′B was able to neutralize respectively 20.7 μg, 20.2 μg and 13.8 μg of
C. d. terrificus venom.</description><identifier>ISSN: 0041-0101</identifier><identifier>EISSN: 1879-3150</identifier><identifier>DOI: 10.1016/0041-0101(89)90177-3</identifier><identifier>PMID: 2728022</identifier><identifier>CODEN: TOXIA6</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Animal poisons toxicology. Antivenoms ; Animals ; Antibody Specificity ; Antivenins - isolation & purification ; Biological and medical sciences ; Blotting, Western ; Caprylates - pharmacology ; Crotalid Venoms - immunology ; Electrophoresis, Polyacrylamide Gel ; Horses - immunology ; Immunoglobulin Fab Fragments - isolation & purification ; Immunoglobulin G - isolation & purification ; Medical sciences ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Neutralization Tests ; Toxicology</subject><ispartof>Toxicon (Oxford), 1989, Vol.27 (3), p.297-303</ispartof><rights>1989</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-1289d62298392b48ec9390510e4cb5104a99bc9955b6f17ca3f6c0288138edcc3</citedby><cites>FETCH-LOGICAL-c386t-1289d62298392b48ec9390510e4cb5104a99bc9955b6f17ca3f6c0288138edcc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0041-0101(89)90177-3$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,4022,27922,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7326855$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2728022$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>dos Santos, M.C.</creatorcontrib><creatorcontrib>D'Império Lima, M.R.</creatorcontrib><creatorcontrib>Furtado, G.C.</creatorcontrib><creatorcontrib>Colletto, G.M.D.D.</creatorcontrib><creatorcontrib>Kipnis, T.L.</creatorcontrib><creatorcontrib>Dias da Silva, W.</creatorcontrib><title>Purification of F(ab′) 2 anti-snake venom by caprylic acid: A fast method for obtaining IgG fragments with high neutralization activity, purity and yield</title><title>Toxicon (Oxford)</title><addtitle>Toxicon</addtitle><description>Pooled horse plasma containing antibodies against
Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab′)
2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab′)
2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab′ fragments were obtained by reduction and alkylation of F(ab′)
2B. The anti-whole
C. d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab′)
2B], 4800 [F(ab′)
2M] and 3200 (Fab′B). Immunochemical analysis of these fragments by SDS gel electrophoresis, Western blot and by double immunodiffusion revealed that the solution containing. F(ab′)
2M was free of IgG and of other plasma proteins, whereas that containing F(ab′)
2B was not. One milligram of either F(ab′)
2B, F(ab′)
2M or Fab′B was able to neutralize respectively 20.7 μg, 20.2 μg and 13.8 μg of
C. d. terrificus venom.</description><subject>Animal poisons toxicology. Antivenoms</subject><subject>Animals</subject><subject>Antibody Specificity</subject><subject>Antivenins - isolation & purification</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Caprylates - pharmacology</subject><subject>Crotalid Venoms - immunology</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Horses - immunology</subject><subject>Immunoglobulin Fab Fragments - isolation & purification</subject><subject>Immunoglobulin G - isolation & purification</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Mice, Inbred C57BL</subject><subject>Neutralization Tests</subject><subject>Toxicology</subject><issn>0041-0101</issn><issn>1879-3150</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1uFDEQhS0ECkPgBiB5gVAi0eCf_rFZIEURCZEiwQLWlrvaPVPQ7R5sz6DOKvfgFhyJk-BhRrNkVSXVV69K7xHynLM3nPH6LWMlL1huz5Q-14w3TSEfkAVXjS4kr9hDsjgij8mTGL8xxqTS9Qk5EY1QTIgF-fV5E7BHsAknT6eeXp3Z9s_973MqqPUJi-jtd0e3zk8jbWcKdh3mAYFawO4dvaC9jYmOLq2mjvZToFObLHr0S3qzvKZ9sMvR-RTpT0wrusLlinq3ScEOeLe_aSHhFtP8mq7zK2nOZzs6oxu6p-RRb4fonh3qKfl69eHL5cfi9tP1zeXFbQFS1angQumuFkIrqUVbKgdaalZx5kpocymt1i1oXVVt3fMGrOxrYEIpLpXrAOQpebXXXYfpx8bFZEaM4IbBejdtomk0k1mxzGC5ByFMMQbXm3XA0YbZcGZ2mZid4WZnuFHa_MvEyLz24qC_aUfXHZcOIeT5y8PcRrBD9swDxiPWSFGrqsrY-z3mshdbdMFEQOfBdRgcJNNN-P8__gL9Bak-</recordid><startdate>1989</startdate><enddate>1989</enddate><creator>dos Santos, M.C.</creator><creator>D'Império Lima, M.R.</creator><creator>Furtado, G.C.</creator><creator>Colletto, G.M.D.D.</creator><creator>Kipnis, T.L.</creator><creator>Dias da Silva, W.</creator><general>Elsevier Ltd</general><general>Elsevier Science</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>1989</creationdate><title>Purification of F(ab′) 2 anti-snake venom by caprylic acid: A fast method for obtaining IgG fragments with high neutralization activity, purity and yield</title><author>dos Santos, M.C. ; D'Império Lima, M.R. ; Furtado, G.C. ; Colletto, G.M.D.D. ; Kipnis, T.L. ; Dias da Silva, W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-1289d62298392b48ec9390510e4cb5104a99bc9955b6f17ca3f6c0288138edcc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Animal poisons toxicology. Antivenoms</topic><topic>Animals</topic><topic>Antibody Specificity</topic><topic>Antivenins - isolation & purification</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Caprylates - pharmacology</topic><topic>Crotalid Venoms - immunology</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Horses - immunology</topic><topic>Immunoglobulin Fab Fragments - isolation & purification</topic><topic>Immunoglobulin G - isolation & purification</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Mice, Inbred C57BL</topic><topic>Neutralization Tests</topic><topic>Toxicology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>dos Santos, M.C.</creatorcontrib><creatorcontrib>D'Império Lima, M.R.</creatorcontrib><creatorcontrib>Furtado, G.C.</creatorcontrib><creatorcontrib>Colletto, G.M.D.D.</creatorcontrib><creatorcontrib>Kipnis, T.L.</creatorcontrib><creatorcontrib>Dias da Silva, W.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Toxicon (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>dos Santos, M.C.</au><au>D'Império Lima, M.R.</au><au>Furtado, G.C.</au><au>Colletto, G.M.D.D.</au><au>Kipnis, T.L.</au><au>Dias da Silva, W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification of F(ab′) 2 anti-snake venom by caprylic acid: A fast method for obtaining IgG fragments with high neutralization activity, purity and yield</atitle><jtitle>Toxicon (Oxford)</jtitle><addtitle>Toxicon</addtitle><date>1989</date><risdate>1989</risdate><volume>27</volume><issue>3</issue><spage>297</spage><epage>303</epage><pages>297-303</pages><issn>0041-0101</issn><eissn>1879-3150</eissn><coden>TOXIA6</coden><abstract>Pooled horse plasma containing antibodies against
Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab′)
2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab′)
2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab′ fragments were obtained by reduction and alkylation of F(ab′)
2B. The anti-whole
C. d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab′)
2B], 4800 [F(ab′)
2M] and 3200 (Fab′B). Immunochemical analysis of these fragments by SDS gel electrophoresis, Western blot and by double immunodiffusion revealed that the solution containing. F(ab′)
2M was free of IgG and of other plasma proteins, whereas that containing F(ab′)
2B was not. One milligram of either F(ab′)
2B, F(ab′)
2M or Fab′B was able to neutralize respectively 20.7 μg, 20.2 μg and 13.8 μg of
C. d. terrificus venom.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>2728022</pmid><doi>10.1016/0041-0101(89)90177-3</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0041-0101 |
| ispartof | Toxicon (Oxford), 1989, Vol.27 (3), p.297-303 |
| issn | 0041-0101 1879-3150 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79033904 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Animal poisons toxicology. Antivenoms Animals Antibody Specificity Antivenins - isolation & purification Biological and medical sciences Blotting, Western Caprylates - pharmacology Crotalid Venoms - immunology Electrophoresis, Polyacrylamide Gel Horses - immunology Immunoglobulin Fab Fragments - isolation & purification Immunoglobulin G - isolation & purification Medical sciences Mice Mice, Inbred BALB C Mice, Inbred C57BL Neutralization Tests Toxicology |
| title | Purification of F(ab′) 2 anti-snake venom by caprylic acid: A fast method for obtaining IgG fragments with high neutralization activity, purity and yield |
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