Development of a double-recombinant antibody sandwich ELISA for quantitative detection of epsilon toxoid concentration in inactivated Clostridium perfringens vaccines

Abstract Background: Epsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals. Also, ETX causes serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, ELISA analysis showed that most isolated clones had high affinity and specificity for ETX.Results: Two recombinant monoclonal antibodies against ETX were isolated by phage display technology. B1 phage VH antibody isolated from DAb library and G2 soluble scFv antibody isolated from Tomlinson I+J libraries have been applied as the capture and detection antibodies for developing an ETX sandwich ELISA test, respectively.Conclusions: Designed ETX sandwich ELISA could be a valuable tool for quantitative detection of ETX in inactivated commercial vaccines against enterotoxemia.

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Development of a double-recombinant antibody sandwich ELISA for quantitative detection of epsilon toxoid concentration in inactivated Clostridium perfringens vaccines

BMC Veterinary Research ◽

10.1186/s12917-020-02572-4 ◽

2020 ◽

Vol 16 (1) ◽

Author(s):

Maryam Alibeiki ◽

Mehdi Golchin ◽

Mohammad Tabatabaei

Keyword(s):

Recombinant Antibody ◽

Sandwich Elisa ◽

Animal Husbandry ◽

Elisa Test ◽

Economic Losses ◽

Quantitative Detection ◽

Phage Display Technology ◽

Epsilon Toxin ◽

Display Technology ◽

Soluble Scfv

Abstract Background Epsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals. Also, ETX causes serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, ELISA analysis showed that most isolated clones had high affinity and specificity for ETX. Results Two recombinant monoclonal antibodies against ETX were isolated by phage display technology. B1 phage VH antibody isolated from DAb library and G2 soluble scFv antibody isolated from Tomlinson I + J libraries have been applied as the capture and detection antibodies for developing an ETX sandwich ELISA test, respectively. Conclusions Designed ETX sandwich ELISA could be a valuable tool for quantitative detection of ETX in inactivated commercial vaccines against enterotoxemia. Graphical abstract

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Development of a double-recombinant antibody sandwich ELISA for quantitative detection of epsilon toxoid concentration in inactivated Clostridium perfringens vaccines

10.21203/rs.3.rs-25433/v3 ◽

2020 ◽

Author(s):

Maryam Alibeiki ◽

Mehdi Golchin ◽

Mohammad Tabatabaei

Keyword(s):

Recombinant Antibody ◽

Sandwich Elisa ◽

Animal Husbandry ◽

Elisa Test ◽

Economic Losses ◽

Quantitative Detection ◽

Phage Display Technology ◽

Epsilon Toxin ◽

Display Technology ◽

Soluble Scfv

Abstract Background: Epsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals. Also, ETX causes serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, ELISA analysis showed that most isolated clones had high affinity and specificity for ETX.Results: Two recombinant monoclonal antibodies against ETX were isolated by phage display technology. B1 phage VH antibody isolated from DAb library and G2 soluble scFv antibody isolated from Tomlinson I+J libraries have been applied as the capture and detection antibodies for developing an ETX sandwich ELISA test, respectively. Conclusions: Designed ETX sandwich ELISA could be a valuable tool for quantitative detection of ETX in inactivated commercial vaccines against enterotoxemia.

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Development of a Double-Recombinant Antibody Sandwich ELISA for Quantitative Detection of Epsilon Toxoid Concentration in Inactivated Clostridium Perfringens Vaccines

10.21203/rs.3.rs-25433/v1 ◽

2020 ◽

Author(s):

Maryam Alibeiki ◽

Mehdi Golchin ◽

Mohammad Tabatabaei

Keyword(s):

Recombinant Antibody ◽

Sandwich Elisa ◽

Animal Husbandry ◽

Elisa Test ◽

Economic Losses ◽

Quantitative Detection ◽

Phage Display Technology ◽

Detection Range ◽

Epsilon Toxin ◽

Display Technology

Abstract Background: Epsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals and causing serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, we evidenced that most isolated clones had high affinity and specificity for the ETX by ELISA.Results: We isolated two recombinant monoclonal antibodies against ETX by phage display technology and Using the B1 phage VH antibody isolated from DAb library as capture antibody and G2 soluble scFv antibody isolated from Tomlinson I + J libraries as the detector antibody, we developed a sandwich ELISA test.Conclusions: This sandwich ELISA could be a good candidate for quantitative detection ETX with the detection range of 5 ~ 100000 ng/ml in inactivated commercial vaccines against enterotoxemia.

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Production of Recombinant Antibody Against Potato Virus Y by using Phage Display Technology

Biotechnology & Biotechnological Equipment ◽

10.1080/13102818.2000.10819056 ◽

2000 ◽

Vol 14 (1) ◽

pp. 18-24

Author(s):

B. Erdag ◽

E. Atalay ◽

B. Çirakoglu

Keyword(s):

Phage Display ◽

Potato Virus ◽

Potato Virus Y ◽

Recombinant Antibody ◽

Phage Display Technology ◽

Display Technology

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Characterization of 11-dehydro-thromboxane B2 recombinant antibody obtained by phage display technology

Prostaglandins Leukotrienes and Essential Fatty Acids ◽

10.1016/s0952-3278(03)00006-1 ◽

2003 ◽

Vol 68 (4) ◽

pp. 273-284 ◽

Cited By ~ 9

Author(s):

Lilian Rumi Tsuruta ◽

Yoshihisa Tomioka ◽

Takanori Hishinuma ◽

Yoshinori Kato ◽

Kunihiko Itoh ◽

...

Keyword(s):

Phage Display ◽

Recombinant Antibody ◽

Thromboxane B2 ◽

Phage Display Technology ◽

Display Technology

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Mimotopes for Mycotoxins Diagnosis Based on Random Peptides or Recombinant Antibodies from Phage Library

Molecules ◽

10.3390/molecules26247652 ◽

2021 ◽

Vol 26 (24) ◽

pp. 7652

Author(s):

Wei Sun ◽

Yan Zhang ◽

Zhigang Ju

Keyword(s):

Phage Display ◽

Recombinant Antibody ◽

Recombinant Antibodies ◽

Phage Display Library ◽

Ease Of Use ◽

Phage Library ◽

Phage Display Technology ◽

Random Peptide ◽

Display Technology

Mycotoxins, the small size secondary metabolites of fungi, have posed a threat to the safety of medicine, food and public health. Therefore, it is essential to create sensitive and effective determination of mycotoxins. Based on the special affinity between antibody and antigen, immunoassay has been proved to be a powerful technology for the detection of small analytes. However, the tedious preparation and instability of conventional antibodies restrict its application on easy and fast mycotoxins detection. By virtue of simplicity, ease of use, and lower cost, phage display library provides novel choices for antibodies or hapten conjugates, and lead random peptide or recombinant antibody to becoming the promising and environmental friendly immune-reagents in the next generation of immunoassays. This review briefly describes the latest developments on mycotoxins detection using M13 phage display, mainly focusing on the recent applications of phage display technology employed in mycotoxins detection, including the introduction of phage and phage display, the types of phage displayed peptide/recombinant antibody library, random peptides/recombinant antibodies-based immunoassays, as well as simultaneous determination of multiple mycotoxins.

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The structure of the C-terminal domain of the nucleoprotein from the Bundibugyo strain of the Ebola virus in complex with a pan-specific synthetic Fab

Acta Crystallographica Section D Structural Biology ◽

10.1107/s2059798318007878 ◽

2018 ◽

Vol 74 (7) ◽

pp. 681-689 ◽

Cited By ~ 3

Author(s):

Malwina J. Radwańska ◽

Mateusz Jaskółowski ◽

Elena Davydova ◽

Urszula Derewenda ◽

Tsuyoshi Miyake ◽

...

Keyword(s):

Phage Display ◽

Ebola Virus ◽

Sandwich Elisa ◽

Phage Display Technology ◽

Synthetic Antibody ◽

Display Technology ◽

Major Bottleneck ◽

Hybridoma Technology ◽

Selection For ◽

Detection Technologies

The vast majority of platforms for the detection of viral or bacterial antigens rely on immunoassays, typically ELISA or sandwich ELISA, that are contingent on the availability of suitable monoclonal antibodies (mAbs). This is a major bottleneck, since the generation and production of mAbs is time-consuming and expensive. Synthetic antibody fragments (sFabs) generated by phage-display selection offer an alternative with many advantages over Fabs obtained from natural antibodies using hybridoma technology. Unlike mAbs, sFabs are generated using phage display, allowing selection for binding to specific strains or for pan-specificity, for identification of structural epitopes or unique protein conformations and even for complexes. Further, they can easily be produced in Escherichia coli in large quantities and engineered for purposes of detection technologies and other applications. Here, the use of phage-display selection to generate a pan-specific Fab (MJ20), based on a Herceptin Fab scaffold, with the ability to bind selectively and with high affinity to the C-terminal domains of the nucleoproteins (NPs) from all five known strains of the Ebola virus is reported. The high-resolution crystal structure of the complex of MJ20 with the antigen from the Bundibugyo strain of the Ebola virus reveals the basis for pan-specificity and illustrates how the phage-display technology can be used to manufacture suitable Fabs for use in diagnostic or therapeutic applications.

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