دورية أكاديمية
Engineered magnetosomes fused to functional molecule (protein A) provide a highly effective alternative to commercial immunomagnetic beads
العنوان: | Engineered magnetosomes fused to functional molecule (protein A) provide a highly effective alternative to commercial immunomagnetic beads |
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المؤلفون: | Junjie Xu, Lingzi Liu, Jinxin He, Shijiao Ma, Shuli Li, Zhanhui Wang, Ting Xu, Wei Jiang, Ying Wen, Ying Li, Jiesheng Tian, Feng Li |
المصدر: | Journal of Nanobiotechnology, Vol 17, Iss 1, Pp 1-11 (2019) |
بيانات النشر: | BMC, 2019. |
سنة النشر: | 2019 |
المجموعة: | LCC:Biotechnology LCC:Medical technology |
مصطلحات موضوعية: | Bacterial magnetic nanoparticles, Protein A, Surface display technique, Gentamicin sulfate, Vibrio parahaemolyticus, Biotechnology, TP248.13-248.65, Medical technology, R855-855.5 |
الوصف: | Abstract Background Magnetosomes (also called bacterial magnetic nanoparticles; BMPs) are biomembrane-coated nanoparticles synthesized by magnetotactic bacteria (MTB). Engineered BMPs fused to protein A (termed ∆F-BMP-FA) bind antibodies (Abs) automatically, and thus provide a series of potential advantages. However, no report so far has systematically evaluated functional applicability of genetically engineered BMPs. Results We evaluated properties of ∆F-BMP-FA, and developed/optimized culture methods for host strain Magnetospirillum gryphiswaldense ΔF-FA, ∆F-BMP-FA extraction conditions, conditions for Ab conjugation to ∆F-BMP-FA surface, and procedures for antigen detection using ∆F-BMP-FA/Ab complexes (termed BMP-A-Ab). Fed-batch culture for 36 h in a 42-L fermentor resulted in yields (dry weight) of 2.26 g/L for strain ΔF-FA and 62 mg/L for ∆F-BMP-FA. Optimal wash cycle number for ∆F-BMP-FA purification was seven, with magnetic separation following each ultrasonication step. Fusion of protein A to BMPs resulted in ordered arrangement of Abs on BMP surface. Linkage rate 962 μg Ab per mg ∆F-BMP-FA was achieved. BMP-A-Ab were tested for detection of pathogen (Vibrio parahaemolyticus; Vp) surface antigen and hapten (gentamicin sulfate). Maximal Vp capture rate for BMP-A-Ab was 90% (higher than rate for commercial immunomagnetic beads), and detection sensitivity was 5 CFU/mL. ∆F-BMP-FA also bound Abs from crude mouse ascites to form complex. Lowest gentamicin sulfate detection line for BMP-A-Ab was 0.01 ng/mL, 400-fold lower than that for double Ab sandwich ELISA, and gentamicin sulfate recovery rate for BMP-A-Ab was 93.2%. Conclusion Our findings indicate that engineered BMPs such as ∆F-BMP-FA are inexpensive, eco-friendly alternatives to commercial immunomagnetic beads for detection or diagnostic immunoassays, and have high Ab-conjugation and antigen-adsorption capacity. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English |
تدمد: | 1477-3155 |
Relation: | http://link.springer.com/article/10.1186/s12951-019-0469-z; https://doaj.org/toc/1477-3155 |
DOI: | 10.1186/s12951-019-0469-z |
URL الوصول: | https://doaj.org/article/f6a8356db71e4cbd91ff708557c1ffa0 |
رقم الأكسشن: | edsdoj.f6a8356db71e4cbd91ff708557c1ffa0 |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 14773155 |
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DOI: | 10.1186/s12951-019-0469-z |