دورية أكاديمية
أعرض في EDS

Nanobodies as potential tools for microbiological testing of live biotherapeutic products.

التفاصيل البيبلوغرافية
العنوان: Nanobodies as potential tools for microbiological testing of live biotherapeutic products.
المؤلفون: Dorosky RJ; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA. Robert.Dorosky@fda.hhs.gov., Schreier JE; Department of Marine Sciences, University of Georgia, Athens, GA, USA., Lola SL; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., Sava RL; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., Coryell MP; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., Akue A; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., KuKuruga M; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., Carlson PE Jr; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., Dreher-Lesnick SM; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA., Stibitz S; Office of Vaccines Research and Review, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.
المصدر: AMB Express [AMB Express] 2024 Jan 20; Vol. 14 (1), pp. 9. Date of Electronic Publication: 2024 Jan 20.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101561785 Publication Model: Electronic Cited Medium: Print ISSN: 2191-0855 (Print) Linking ISSN: 21910855 NLM ISO Abbreviation: AMB Express Subsets: PubMed not MEDLINE
أسماء مطبوعة: Original Publication: Heidelberg : Springer-Verlag, c2011-
مستخلص: Nanobodies are highly specific binding domains derived from naturally occurring single chain camelid antibodies. Live biotherapeutic products (LBPs) are biological products containing preparations of live organisms, such as Lactobacillus, that are intended for use as drugs, i.e. to address a specific disease or condition. Demonstrating potency of multi-strain LBPs can be challenging. The approach investigated here is to use strain-specific nanobody reagents in LBP potency assays. Llamas were immunized with radiation-killed Lactobacillus jensenii or L. crispatus whole cell preparations. A nanobody phage-display library was constructed and panned against bacterial preparations to identify nanobodies specific for each species. Nanobody-encoding DNA sequences were subcloned and the nanobodies were expressed, purified, and characterized. Colony immunoblots and flow cytometry showed that binding by Lj75 and Lj94 nanobodies were limited to a subset of L. jensenii strains while binding by Lc38 and Lc58 nanobodies were limited to L. crispatus strains. Mass spectrometry was used to demonstrate that Lj75 specifically bound a peptidase of L. jensenii, and that Lc58 bound an S-layer protein of L. crispatus. The utility of fluorescent nanobodies in evaluating multi-strain LBP potency assays was assessed by evaluating a L. crispatus and L. jensenii mixture by fluorescence microscopy, flow cytometry, and colony immunoblots. Our results showed that the fluorescent nanobody labelling enabled differentiation and quantitation of the strains in mixture by these methods. Development of these nanobody reagents represents a potential advance in LBP testing, informing the advancement of future LBP potency assays and, thereby, facilitation of clinical investigation of LBPs.
(© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
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معلومات مُعتمدة: AAI17016-001-00001 Division of Intramural Research, National Institute of Allergy and Infectious Diseases
فهرسة مساهمة: Keywords: Biologics; Lactobacillus; Live biotherapeutic products; Nanobody; Potency assays
تواريخ الأحداث: Date Created: 20240120 Latest Revision: 20240123
رمز التحديث: 20240123
مُعرف محوري في PubMed: PMC10799837
DOI: 10.1186/s13568-023-01659-z
PMID: 38245586
قاعدة البيانات: MEDLINE
الوصف
تدمد:2191-0855
DOI:10.1186/s13568-023-01659-z