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Construction of an immunized rabbit phage display antibody library for screening microcystin-LR high sensitive single-chain antibody
- Source :
- International journal of biological macromolecules. 123
- Publication Year :
- 2018
-
Abstract
- Microcystin-LR (MC-LR) is one of the most common biotoxin that pollutes water and agricultural products. The study aims to obtain the high sensitive anti-MC-LR single-chain antibody (scFv) for detecting MC-LR. Here, a MC-LR-immunized rabbit phage display scFv library with its capacity of 3.26×109CFU/mL was constructed and used for anti-MC-LR phage scFv particles screening. After four rounds of biopanning, 18 positives were isolated and identified successfully. The most positive scFv (RscFv3) was expressed in Escherichia coli HB2151 and purified with a concentration of 796.7μg/mL, and the purified anti-MC-LR polyclonal antibodies (PAbs) were 3.6mg/mL. The PAbs and scFv based indirect competitive enzyme linked immunosorbent assay (IC-ELISAs) were established for MC-LR and its analogues, and the results showed they all had high sensitive for MC-LR with detection limits of 0.03 and 0.05μg/L, and had strong cross-reactivity for MC-RR, MC-WR and MC-YR, respectively. The average recovery rate was 91.9% with coefficient of variation
- Subjects :
- Phage display
Microcystins
chemical and pharmacologic phenomena
Microcystin-LR
02 engineering and technology
Biopanning
medicine.disease_cause
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Structural Biology
Antibody Specificity
Peptide Library
medicine
Animals
Molecular Biology
Escherichia coli
030304 developmental biology
chemistry.chemical_classification
Detection limit
0303 health sciences
biology
General Medicine
respiratory system
021001 nanoscience & nanotechnology
Molecular biology
Enzyme
chemistry
Polyclonal antibodies
biology.protein
Marine Toxins
Rabbits
Antibody
0210 nano-technology
Single-Chain Antibodies
Subjects
Details
- ISSN :
- 18790003
- Volume :
- 123
- Database :
- OpenAIRE
- Journal :
- International journal of biological macromolecules
- Accession number :
- edsair.doi.dedup.....7a2d06acb7adf189fea5b34174a1c94c