An alternate method utilizing small quantities of ligand for affinity purification of monospecific antibodies
An alternate method was designed to couple a limited quantity of protein to an affinity support when a conventional technique was unsuccessful. This was achieved through the introduction of a small number of sulfhydryl groups to the ligand by reaction with 2-iminothiolane which resulted in a limited...
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| Veröffentlicht in: | Journal of immunological methods 1988-08, Vol.112 (1), p.63-69 |
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| container_end_page | 69 |
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| container_issue | 1 |
| container_start_page | 63 |
| container_title | Journal of immunological methods |
| container_volume | 112 |
| creator | Aithal, H.Naga Knigge, Kevin M. Kartha, Sreedharan Czyzewski, E.Ann Toback, F.Gary |
| description | An alternate method was designed to couple a limited quantity of protein to an affinity support when a conventional technique was unsuccessful. This was achieved through the introduction of a small number of sulfhydryl groups to the ligand by reaction with 2-iminothiolane which resulted in a limited number of reactive sites on the protein. Amino groups on an AH-Sepharose 4B matrix were linked to sulfhydryl groups on the ligand using the heterobifunctional agent
m-maleimidobenzoyl sulfosuccinimide ester (sulfo-MBS). This method was employed to prepare an affinity support using a cytosolic protein that actives glyceraldehyde-3-phosphate dehydrogenase as a ligand. Monospecific antibody purified from the affinity column recognized only this protein on a Western blot of a cytosolic extract of kidney epithelial cells. |
| doi_str_mv | 10.1016/0022-1759(88)90034-8 |
| format | Article |
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m-maleimidobenzoyl sulfosuccinimide ester (sulfo-MBS). This method was employed to prepare an affinity support using a cytosolic protein that actives glyceraldehyde-3-phosphate dehydrogenase as a ligand. Monospecific antibody purified from the affinity column recognized only this protein on a Western blot of a cytosolic extract of kidney epithelial cells.</description><subject>Affinity chromatography</subject><subject>Animals</subject><subject>Antibodies, Monoclonal - isolation & purification</subject><subject>Antibody Specificity</subject><subject>Binding Sites, Antibody</subject><subject>Biological and medical sciences</subject><subject>Chromatography, Affinity - methods</subject><subject>Cross-Linking Reagents</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Glyceraldehyde-3-Phosphate Dehydrogenases</subject><subject>Humans</subject><subject>Immunochemical method</subject><subject>Immunoglobulin</subject><subject>Immunoglobulin G - isolation & purification</subject><subject>Immunoglobulin G - metabolism</subject><subject>Immunohistochemistry - methods</subject><subject>Protein modification</subject><subject>Rabbits</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV9rFTEQxYNY6rX6DRTyoujDarKbvy-FUtQWCr7oc8gmkxrZTW6TrFA_vbveS32zTwMzv3OYOYPQK0o-UELFR0L6vqOS63dKvdeEDKxTT9COKtl3UhP-FO0ekGfoea0_CSGUCHKKTgdGBq3JDs0XCdupQUm2AZ6h_cgeLy1O8XdMt7jOdprw3WJTiy1CxTngKd7a5HHIBdsQYortHu-XEkN0tsWcNmbOKdc9uK2JN_GY_Sp_gU6CnSq8PNYz9P3zp2-XV93N1y_Xlxc3nWNUtY5TkJYLEfTAeuaDoL0MQkptPQ3KSQWUa0ld8FopSxjXhOqx70cO662jHM7Q24PvvuS7BWozc6wOpskmyEs1Ug1CcMUfBSknomearSA7gK7kWgsEsy9xtuXeUGK2d5gta7NlbZQyf99h1Cp7ffRfxhn8g-iY_zp_c5zb6uwUik0u1n_emgvF1XbQ-YGDNbVfEYqpLkJy4GMB14zP8f-L_AGgSKdA</recordid><startdate>19880809</startdate><enddate>19880809</enddate><creator>Aithal, H.Naga</creator><creator>Knigge, Kevin M.</creator><creator>Kartha, Sreedharan</creator><creator>Czyzewski, E.Ann</creator><creator>Toback, F.Gary</creator><general>Elsevier B.V</general><general>Elsevier</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>7T5</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19880809</creationdate><title>An alternate method utilizing small quantities of ligand for affinity purification of monospecific antibodies</title><author>Aithal, H.Naga ; Knigge, Kevin M. ; Kartha, Sreedharan ; Czyzewski, E.Ann ; Toback, F.Gary</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c418t-51e7a566f93424df6127f6779ad1f8c78e15971cfd988a0459019b22b5e759b73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Affinity chromatography</topic><topic>Animals</topic><topic>Antibodies, Monoclonal - isolation & purification</topic><topic>Antibody Specificity</topic><topic>Binding Sites, Antibody</topic><topic>Biological and medical sciences</topic><topic>Chromatography, Affinity - methods</topic><topic>Cross-Linking Reagents</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Glyceraldehyde-3-Phosphate Dehydrogenases</topic><topic>Humans</topic><topic>Immunochemical method</topic><topic>Immunoglobulin</topic><topic>Immunoglobulin G - isolation & purification</topic><topic>Immunoglobulin G - metabolism</topic><topic>Immunohistochemistry - methods</topic><topic>Protein modification</topic><topic>Rabbits</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Aithal, H.Naga</creatorcontrib><creatorcontrib>Knigge, Kevin M.</creatorcontrib><creatorcontrib>Kartha, Sreedharan</creatorcontrib><creatorcontrib>Czyzewski, E.Ann</creatorcontrib><creatorcontrib>Toback, F.Gary</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>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Aithal, H.Naga</au><au>Knigge, Kevin M.</au><au>Kartha, Sreedharan</au><au>Czyzewski, E.Ann</au><au>Toback, F.Gary</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An alternate method utilizing small quantities of ligand for affinity purification of monospecific antibodies</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>1988-08-09</date><risdate>1988</risdate><volume>112</volume><issue>1</issue><spage>63</spage><epage>69</epage><pages>63-69</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>An alternate method was designed to couple a limited quantity of protein to an affinity support when a conventional technique was unsuccessful. 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m-maleimidobenzoyl sulfosuccinimide ester (sulfo-MBS). This method was employed to prepare an affinity support using a cytosolic protein that actives glyceraldehyde-3-phosphate dehydrogenase as a ligand. Monospecific antibody purified from the affinity column recognized only this protein on a Western blot of a cytosolic extract of kidney epithelial cells.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>3403990</pmid><doi>10.1016/0022-1759(88)90034-8</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-1759 |
| ispartof | Journal of immunological methods, 1988-08, Vol.112 (1), p.63-69 |
| issn | 0022-1759 1872-7905 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Affinity chromatography Animals Antibodies, Monoclonal - isolation & purification Antibody Specificity Binding Sites, Antibody Biological and medical sciences Chromatography, Affinity - methods Cross-Linking Reagents Female Fundamental and applied biological sciences. Psychology Fundamental immunology Glyceraldehyde-3-Phosphate Dehydrogenases Humans Immunochemical method Immunoglobulin Immunoglobulin G - isolation & purification Immunoglobulin G - metabolism Immunohistochemistry - methods Protein modification Rabbits |
| title | An alternate method utilizing small quantities of ligand for affinity purification of monospecific antibodies |
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