{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T04:05:40Z","timestamp":1784174740543,"version":"3.55.0"},"reference-count":47,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T00:00:00Z","timestamp":1784073600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T00:00:00Z","timestamp":1784073600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["aiche.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotechnology Progress"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Development of therapeutic bispecific antibodies (BsAbs) poses significant manufacturability challenges associated with correct chain pairing and an increased diversity of impurities. Here we address this with a product quality assessment during cell line development (CLD), forgoing the need for genetic characterization during cell line screening. Using Chinese Hamster Ovary (CHO) cell lines expressing a panel of monoclonal antibodies (mAb) and BsAbs, we demonstrate the highly specific binding of SpotLight\u2122 Hu\u03bb, Hu\u0138 and HuFc reagents to their target chain isotypes (lambda or kappa light chain and Fc regions) during single cell cloning workflows on a Bruker Beacon\u00ae Optofluidic System. We then demonstrate combinatorial use of SpotLight\u2122 Hu\u03bb and Hu\u0138 reagents as an early product quality indicator on a CHO cell line expressing a BsAb with dual kappa\u2010lambda light chain configuration. Cell lines presenting equal binding profiles for each SpotLight assay exhibited platform\u2010typical heterodimer levels with an average of 87.26% following fed\u2010batch bioreactor production assessment. Conversely, cell lines biased towards either SpotLight\u2122 assay resulted in lower than desired heterodimer (0%\u201384.23%) and increased associated impurities. This study demonstrates the application of dual SpotLight\u2122 assays as an identifier of CHO cell lines expressing a kappa\u2010lambda light chain BsAb with a high heterodimer percentage. This allows cell lines with optimal product quality to be identified earlier in CLD, resulting in fewer cell lines being progressed to resource\u2010intensive fed\u2010batch studies and subsequent product quality assessment. Furthermore, this assay provides an opportunity to characterize impurity expression biases prior to developing downstream purification processes.<\/jats:p>","DOI":"10.1002\/btpr.88539","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T04:01:39Z","timestamp":1784174499000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Screening of bispecific antibodies during single cell cloning of recombinant Chinese hamster ovary cell lines to improve titer and product quality"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3642-3818","authenticated-orcid":false,"given":"Molly","family":"Robinson","sequence":"first","affiliation":[{"name":"BioPharm Process Research, Drug Substance Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3972-7699","authenticated-orcid":false,"given":"Jessica","family":"Willmott","sequence":"additional","affiliation":[{"name":"BioPharm Process Research, Drug Substance Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holly","family":"Corrigall","sequence":"additional","affiliation":[{"name":"BioPharm Process Research, Drug Substance Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Lega","sequence":"additional","affiliation":[{"name":"BioPharm Process Research, Drug Substance Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-3934-1609","authenticated-orcid":false,"given":"Joseph","family":"Stirling","sequence":"additional","affiliation":[{"name":"BioPharm Process Research, Drug Substance Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8301-614X","authenticated-orcid":false,"given":"Vithiya","family":"Vimalraj","sequence":"additional","affiliation":[{"name":"Drug Substance and Product Analysis, Analytical Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitul","family":"Patel","sequence":"additional","affiliation":[{"name":"Drug Substance and Product Analysis, Analytical Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7521-4335","authenticated-orcid":false,"given":"Robyn","family":"Emmins","sequence":"additional","affiliation":[{"name":"BioPharm Process Research, Drug Substance Development GSK  Stevenage UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,15]]},"reference":[{"issue":"12","key":"e_1_2_11_2_1","doi-asserted-by":"crossref","first-page":"1722","DOI":"10.1038\/s41587-022-01582-x","article-title":"Biopharmaceutical benchmarks 2022","volume":"40","author":"Walsh G","year":"2022","journal-title":"Nat Biotechnol"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41571\u2010024\u201000905\u2010y"},{"issue":"9","key":"e_1_2_11_4_1","doi-asserted-by":"crossref","first-page":"3583","DOI":"10.1016\/j.apsb.2023.05.023","article-title":"Bispecific antibodies in cancer therapy: target selection and regulatory requirements","volume":"13","author":"Sun Y","year":"2023","journal-title":"Acta Pharm Sin B"},{"issue":"1","key":"e_1_2_11_5_1","doi-asserted-by":"crossref","DOI":"10.1155\/2024\/1804038","article-title":"The potential of single\u2010chain variable fragment antibody: role in future therapeutic and 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