Characterization of Anti-Citrinin Specific ScFvs Selected from Non-Immunized Mouse Splenocytes by Eukaryotic Ribosome Display

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2015 Jul 2

PMID 26131718

Citations 5

Authors

Haiwei Cheng

Yifei Chen

Yi Yang

Xueqiu Chen

Xiaolu Guo

Aifang Du

Affiliations

Soon will be listed here.

Abstract

Single chain variable fragments (scFvs) against citrinin (CIT) were selected from a scFv library constructed from the splenocytes of non-immunized mice by an improved eukaryotic ribosome display technology in this study. Bovine serum albumin (BSA)/ CIT-BSA and ovalbumin (OVA)/ CIT-OVA were used as the antigens to select specific anti-CIT scFvs. Eukaryotic in situ RT-PCR method was used to recover the selected mRNA after every affinity selection. After six rounds of ribosome display, expression vector pTIG-TRX carrying specific scFv DNAs were constructed and transformed into Escherichia coli BL21 (DE3) for protein expression. Thirteen positive clones were selected out of which three (designated 23, 68 and 109) showed high binding activity and specificity to CIT by indirect ELISA, while no clone showed binding activity with carrier proteins. The three scFvs showed high specificity to CIT and the cross reactivity with other mycotoxins was below 0.01% as determined by indirect competitive ELISA. These specific scFvs offer a potential novel immunoassay method for CIT residues. This study confirmed the effectiveness of the improved eukaryotic ribosome display system and could be used as a reference for the selection of scFvs specific to other small molecules using ribosome display.

Citing Articles

Recent Progress in Rapid Determination of Mycotoxins Based on Emerging Biorecognition Molecules: A Review.

Wang Y, Zhang C, Wang J, Knopp D Toxins (Basel). 2022; 14(2).

PMID: 35202100 PMC: 8874725. DOI: 10.3390/toxins14020073.

Recombinant antibodies and their use for food immunoanalysis.

Peltomaa R, Barderas R, Benito-Pena E, Moreno-Bondi M Anal Bioanal Chem. 2021; 414(1):193-217.

PMID: 34417836 PMC: 8380008. DOI: 10.1007/s00216-021-03619-7.

Development of Real-Time Immuno-PCR Based on Phage Displayed an Anti-Idiotypic Nanobody for Quantitative Determination of Citrinin in .

Huang W, Tu Z, Ning Z, He Q, Li Y Toxins (Basel). 2019; 11(10).

PMID: 31575068 PMC: 6832940. DOI: 10.3390/toxins11100572.

Ribosome Display: A Potent Display Technology used for Selecting and Evolving Specific Binders with Desired Properties.

Li R, Kang G, Hu M, Huang H Mol Biotechnol. 2018; 61(1):60-71.

PMID: 30406440 DOI: 10.1007/s12033-018-0133-0.

The Preparation and Identification of a Monoclonal Antibody against Citrinin and the Development of Detection via Indirect Competitive ELISA.

Yirga S, Ling S, Yang Y, Yuan J, Wang S Toxins (Basel). 2017; 9(3).

PMID: 28304351 PMC: 5371865. DOI: 10.3390/toxins9030110.

References

Rothe A, Nathanielsz A, Oberhauser F, von Pogge S, Engert A, Hudson P . Ribosome display and selection of human anti-cD22 scFvs derived from an acute lymphocytic leukemia patient. Biol Chem. 2008; 389(4):433-9. DOI: 10.1515/BC.2008.045. View

Qi Y, Wu C, Zhang S, Wang Z, Huang S, Dai L . Selection of anti-sulfadimidine specific ScFvs from a hybridoma cell by eukaryotic ribosome display. PLoS One. 2009; 4(7):e6427. PMC: 2712767. DOI: 10.1371/journal.pone.0006427. View

Yang L, Ding H, Gu Z, Zhao J, Chen H, Tian F . Selection of single chain fragment variables with direct coating of aflatoxin B1 to enzyme-linked immunosorbent assay plates. J Agric Food Chem. 2009; 57(19):8927-32. DOI: 10.1021/jf9019536. View

Li F, Su P, Lin C, Li H, Cheng J, Shi D . Ribosome display and selection of human anti-placental growth factor scFv derived from ovarian cancer patients. Protein Pept Lett. 2009; 17(5):585-90. DOI: 10.2174/092986610791112729. View

Sun Y, Ning B, Liu M, Gao X, Fan X, Liu J . Selection of diethylstilbestrol-specific single-chain antibodies from a non-immunized mouse ribosome display library. PLoS One. 2012; 7(3):e33186. PMC: 3302827. DOI: 10.1371/journal.pone.0033186. View

Tuomi T, JOHNSSON T, Hintikka E, Reijula K . Detection of aflatoxins (G(1-2), B(1-2)), sterigmatocystin, citrinine and ochratoxin A in samples contaminated by microbes. Analyst. 2001; 126(9):1545-50. DOI: 10.1039/b103790k. View

Abramson D, Usleber E, Martlbauer E . Rapid determination of citrinin in corn by fluorescence liquid chromatography and enzyme immunoassay. J AOAC Int. 1999; 82(6):1353-6. View

Reinhard H, Zimmerli B . Reversed-phase liquid chromatographic behavior of the mycotoxins citrinin and ochratoxin A. J Chromatogr A. 1999; 862(2):147-59. DOI: 10.1016/s0021-9673(99)00929-2. View

He M, Menges M, Groves M, Corps E, Liu H, Bruggemann M . Selection of a human anti-progesterone antibody fragment from a transgenic mouse library by ARM ribosome display. J Immunol Methods. 2000; 231(1-2):105-17. DOI: 10.1016/s0022-1759(99)00144-1. View

10.

Hanes J, Schaffitzel C, Knappik A, Pluckthun A . Picomolar affinity antibodies from a fully synthetic naive library selected and evolved by ribosome display. Nat Biotechnol. 2000; 18(12):1287-92. DOI: 10.1038/82407. View

11.

Jermutus L, Honegger A, Schwesinger F, Hanes J, Pluckthun A . Tailoring in vitro evolution for protein affinity or stability. Proc Natl Acad Sci U S A. 2001; 98(1):75-80. PMC: 14547. DOI: 10.1073/pnas.98.1.75. View

12.

Moghaddam A, Lobersli I, Gebhardt K, Braunagel M, Marvik O . Selection and characterisation of recombinant single-chain antibodies to the hapten Aflatoxin-B1 from naive recombinant antibody libraries. J Immunol Methods. 2001; 254(1-2):169-81. DOI: 10.1016/s0022-1759(01)00413-6. View

13.

Lamla T, Erdmann V . In vitro selection of other proteins than antibodies by means of ribosome display. FEBS Lett. 2001; 502(1-2):35-40. DOI: 10.1016/s0014-5793(01)02660-6. View

14.

Charlton K, Harris W, Porter A . The isolation of super-sensitive anti-hapten antibodies from combinatorial antibody libraries derived from sheep. Biosens Bioelectron. 2001; 16(9-12):639-46. DOI: 10.1016/s0956-5663(01)00192-0. View

15.

Zhou J, Fujita S, Warashina M, Baba T, Taira K . A novel strategy by the action of ricin that connects phenotype and genotype without loss of the diversity of libraries. J Am Chem Soc. 2002; 124(4):538-43. DOI: 10.1021/ja011885x. View

16.

Takahashi F, Ebihara T, Mie M, Yanagida Y, Endo Y, Kobatake E . Ribosome display for selection of active dihydrofolate reductase mutants using immobilized methotrexate on agarose beads. FEBS Lett. 2002; 514(1):106-10. DOI: 10.1016/s0014-5793(02)02334-7. View

17.

Lin C . Simple and sensitive determination of citrinin in Monascus by GC-selected ion monitoring mass spectrometry. Anal Sci. 2002; 18(3):283-7. DOI: 10.2116/analsci.18.283. View

18.

Amstutz P, Pelletier J, Guggisberg A, Jermutus L, Cesaro-Tadic S, Zahnd C . In vitro selection for catalytic activity with ribosome display. J Am Chem Soc. 2002; 124(32):9396-403. DOI: 10.1021/ja025870q. View

19.

Lamla T, Erdmann V . Searching sequence space for high-affinity binding peptides using ribosome display. J Mol Biol. 2003; 329(2):381-8. DOI: 10.1016/s0022-2836(03)00432-7. View

20.

Bennett J, Klich M . Mycotoxins. Clin Microbiol Rev. 2003; 16(3):497-516. PMC: 164220. DOI: 10.1128/CMR.16.3.497-516.2003. View