Analysis of the in vitro function and internalization ability of a humanized EGFR antibody AE01 expressed by Chinese hamster ovary cells
The primary objective of this study was to obtain humanized EGFR antibody and to study it in vitro binding and endocytosis to A431 epidermoid carcinoma cells overexpressing EGFR. Firstly, humanized anti-EGFR AE01 was stably expressed in CHO system. The expression of AE01 was detected by SDS-PAGE and...
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| Veröffentlicht in: | Protein expression and purification 2023-06, Vol.206, p.106243-106243, Article 106243 |
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| container_title | Protein expression and purification |
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| creator | Wu, Hao Ding, Xuefeng Chen, Yun Cai, Yanfei Yang, Zhaoqi Xu, Qinghe Jin, Jian |
| description | The primary objective of this study was to obtain humanized EGFR antibody and to study it in vitro binding and endocytosis to A431 epidermoid carcinoma cells overexpressing EGFR. Firstly, humanized anti-EGFR AE01 was stably expressed in CHO system. The expression of AE01 was detected by SDS-PAGE and Western blot. The binding and endocytosis of AE01 were detected by flow cytometry and immunofluorescence assay. The results showed that: (1) Pure humanized AE01 was prepared, (2) AE01 specifically binds to A431 cells on the cell surface (EGFR-positive), but not binds to NIH 3T3 cells (EGFR-negative), (3) AE01 can effectively inhibit the proliferation of A431 cells, and (4) AE01 binds to A431 cell surface triggered internalization. The antibody is expected to be a candidate molecule for EGFR overexpressed cancer cell targeted therapeutic vectors.
•First, full-length humanized EGFR antibodies (AE01) were successfully prepared from Chinese hamster ovary cells.•Second, AE01 can specifically recognize A431 cells with high EGFR expression.•Finally, AE01 binds to A431 cell surface triggered internalization. |
| doi_str_mv | 10.1016/j.pep.2023.106243 |
| format | Article |
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•First, full-length humanized EGFR antibodies (AE01) were successfully prepared from Chinese hamster ovary cells.•Second, AE01 can specifically recognize A431 cells with high EGFR expression.•Finally, AE01 binds to A431 cell surface triggered internalization.</description><identifier>ISSN: 1046-5928</identifier><identifier>EISSN: 1096-0279</identifier><identifier>DOI: 10.1016/j.pep.2023.106243</identifier><identifier>PMID: 36754125</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Antibodies ; Antibody ; Cell Line, Tumor ; Chinese hamster ovary cells ; CHO Cells ; Cricetinae ; Cricetulus ; Epidermal growth factor receptor ; ErbB Receptors ; Expression ; Humanized ; Internalization ; Mice</subject><ispartof>Protein expression and purification, 2023-06, Vol.206, p.106243-106243, Article 106243</ispartof><rights>2023 Elsevier Inc.</rights><rights>Copyright © 2023 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c353t-c83f666537897ae983cd746fac69c0c9b72666e329d858f6d836be2f642da8e93</citedby><cites>FETCH-LOGICAL-c353t-c83f666537897ae983cd746fac69c0c9b72666e329d858f6d836be2f642da8e93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.pep.2023.106243$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36754125$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wu, Hao</creatorcontrib><creatorcontrib>Ding, Xuefeng</creatorcontrib><creatorcontrib>Chen, Yun</creatorcontrib><creatorcontrib>Cai, Yanfei</creatorcontrib><creatorcontrib>Yang, Zhaoqi</creatorcontrib><creatorcontrib>Xu, Qinghe</creatorcontrib><creatorcontrib>Jin, Jian</creatorcontrib><title>Analysis of the in vitro function and internalization ability of a humanized EGFR antibody AE01 expressed by Chinese hamster ovary cells</title><title>Protein expression and purification</title><addtitle>Protein Expr Purif</addtitle><description>The primary objective of this study was to obtain humanized EGFR antibody and to study it in vitro binding and endocytosis to A431 epidermoid carcinoma cells overexpressing EGFR. Firstly, humanized anti-EGFR AE01 was stably expressed in CHO system. The expression of AE01 was detected by SDS-PAGE and Western blot. The binding and endocytosis of AE01 were detected by flow cytometry and immunofluorescence assay. The results showed that: (1) Pure humanized AE01 was prepared, (2) AE01 specifically binds to A431 cells on the cell surface (EGFR-positive), but not binds to NIH 3T3 cells (EGFR-negative), (3) AE01 can effectively inhibit the proliferation of A431 cells, and (4) AE01 binds to A431 cell surface triggered internalization. The antibody is expected to be a candidate molecule for EGFR overexpressed cancer cell targeted therapeutic vectors.
•First, full-length humanized EGFR antibodies (AE01) were successfully prepared from Chinese hamster ovary cells.•Second, AE01 can specifically recognize A431 cells with high EGFR expression.•Finally, AE01 binds to A431 cell surface triggered internalization.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Antibody</subject><subject>Cell Line, Tumor</subject><subject>Chinese hamster ovary cells</subject><subject>CHO Cells</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>Epidermal growth factor receptor</subject><subject>ErbB Receptors</subject><subject>Expression</subject><subject>Humanized</subject><subject>Internalization</subject><subject>Mice</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1u1DAUhS0EoqXwAGyQl2wy9U9ix2I1Gk0LUiUkBGvLsW80HiVxsJMR6RP0sXGUwrIr2_d-5-heH4Q-UrKjhIrb826EcccI4_ktWMlfoWtKlCgIk-r1ei9FUSlWX6F3KZ0JoVSQ6i264kJWJWXVNXraD6Zbkk84tHg6AfYDvvgpBtzOg518GLAZXK5OEDPpH81Wa3znp2UVGXyaezP4R3D4eH_3I_OTb4Jb8P5IKIY_Y4SUcrNZ8OHkB0iAT6ZP2RCHi4kLttB16T1605ouwYfn8wb9ujv-PHwtHr7ffzvsHwrLKz4VtuatEKLislbSgKq5dbIUrbFCWWJVI1luA2fK1VXdCldz0QBrRcmcqUHxG_R58x1j-D1DmnTv0zqBGSDMSTMpy2wtK5pRuqE2hpQitHqMvs8Ta0r0GoA-6xyAXgPQWwBZ8-nZfm56cP8V_348A182APKSFw9RJ-thsOB8BDtpF_wL9n8Bg32XIg</recordid><startdate>202306</startdate><enddate>202306</enddate><creator>Wu, Hao</creator><creator>Ding, Xuefeng</creator><creator>Chen, Yun</creator><creator>Cai, Yanfei</creator><creator>Yang, Zhaoqi</creator><creator>Xu, Qinghe</creator><creator>Jin, Jian</creator><general>Elsevier Inc</general><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>202306</creationdate><title>Analysis of the in vitro function and internalization ability of a humanized EGFR antibody AE01 expressed by Chinese hamster ovary cells</title><author>Wu, Hao ; Ding, Xuefeng ; Chen, Yun ; Cai, Yanfei ; Yang, Zhaoqi ; Xu, Qinghe ; Jin, Jian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c353t-c83f666537897ae983cd746fac69c0c9b72666e329d858f6d836be2f642da8e93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Antibody</topic><topic>Cell Line, Tumor</topic><topic>Chinese hamster ovary cells</topic><topic>CHO Cells</topic><topic>Cricetinae</topic><topic>Cricetulus</topic><topic>Epidermal growth factor receptor</topic><topic>ErbB Receptors</topic><topic>Expression</topic><topic>Humanized</topic><topic>Internalization</topic><topic>Mice</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wu, Hao</creatorcontrib><creatorcontrib>Ding, Xuefeng</creatorcontrib><creatorcontrib>Chen, Yun</creatorcontrib><creatorcontrib>Cai, Yanfei</creatorcontrib><creatorcontrib>Yang, Zhaoqi</creatorcontrib><creatorcontrib>Xu, Qinghe</creatorcontrib><creatorcontrib>Jin, Jian</creatorcontrib><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>Protein expression and purification</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wu, Hao</au><au>Ding, Xuefeng</au><au>Chen, Yun</au><au>Cai, Yanfei</au><au>Yang, Zhaoqi</au><au>Xu, Qinghe</au><au>Jin, Jian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Analysis of the in vitro function and internalization ability of a humanized EGFR antibody AE01 expressed by Chinese hamster ovary cells</atitle><jtitle>Protein expression and purification</jtitle><addtitle>Protein Expr Purif</addtitle><date>2023-06</date><risdate>2023</risdate><volume>206</volume><spage>106243</spage><epage>106243</epage><pages>106243-106243</pages><artnum>106243</artnum><issn>1046-5928</issn><eissn>1096-0279</eissn><abstract>The primary objective of this study was to obtain humanized EGFR antibody and to study it in vitro binding and endocytosis to A431 epidermoid carcinoma cells overexpressing EGFR. Firstly, humanized anti-EGFR AE01 was stably expressed in CHO system. The expression of AE01 was detected by SDS-PAGE and Western blot. The binding and endocytosis of AE01 were detected by flow cytometry and immunofluorescence assay. The results showed that: (1) Pure humanized AE01 was prepared, (2) AE01 specifically binds to A431 cells on the cell surface (EGFR-positive), but not binds to NIH 3T3 cells (EGFR-negative), (3) AE01 can effectively inhibit the proliferation of A431 cells, and (4) AE01 binds to A431 cell surface triggered internalization. The antibody is expected to be a candidate molecule for EGFR overexpressed cancer cell targeted therapeutic vectors.
•First, full-length humanized EGFR antibodies (AE01) were successfully prepared from Chinese hamster ovary cells.•Second, AE01 can specifically recognize A431 cells with high EGFR expression.•Finally, AE01 binds to A431 cell surface triggered internalization.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>36754125</pmid><doi>10.1016/j.pep.2023.106243</doi><tpages>1</tpages></addata></record> |
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| subjects | Animals Antibodies Antibody Cell Line, Tumor Chinese hamster ovary cells CHO Cells Cricetinae Cricetulus Epidermal growth factor receptor ErbB Receptors Expression Humanized Internalization Mice |
| title | Analysis of the in vitro function and internalization ability of a humanized EGFR antibody AE01 expressed by Chinese hamster ovary cells |
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