Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX

Wang, Rongzhi; Huang, Ailing; Liu, Licai; Xiang, Shuangshuang; Li, Xiufeng; Ling, Sumei; Wang, Lei; Lu, Tun; Wang, Shihua

doi:10.1016/j.toxicon.2014.02.021

Cited by 36 publications

(38 citation statements)

References 57 publications

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“…This scFv was constructed to detect TLH directly in real samples. Furthermore, an antibody was developed against TLH that showed strong neutralizing effects on TLH-induced cell toxicity (Wang et al, , 2007(Wang et al, , 2014aChen et al, 2015).…”

Section: Single-chain Variable Fragment (Scfv)mentioning

confidence: 99%

“…Another technique for recombinant antibody production is the utilization of phage recombinants displaying antibodies at their tips, and which undergo biopanning for the in vitro selection of scFv from large libraries of variable domains circumventing the traditional hybridoma method. Numerous scFv fragments have been constructed against haptens (Wang et al, , 2014a, proteins, carbohydrates, receptors, and tumor antigens for medical therapies and diagnostic applications .…”

Section: Library Construction Of Scfvmentioning

confidence: 99%

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Antibody Engineering for Pursuing a Healthier Future

et al. 2017

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Since the development of antibody-production techniques, a number of immunoglobulins have been developed on a large scale using conventional methods. Hybridoma technology opened a new horizon in the production of antibodies against target antigens of infectious pathogens, malignant diseases including autoimmune disorders, and numerous potent toxins. However, these clinical humanized or chimeric murine antibodies have several limitations and complexities. Therefore, to overcome these difficulties, recent advances in genetic engineering techniques and phage display technique have allowed the production of highly specific recombinant antibodies. These engineered antibodies have been constructed in the hunt for novel therapeutic drugs equipped with enhanced immunoprotective abilities, such as engaging immune effector functions, effective development of fusion proteins, efficient tumor and tissue penetration, and high-affinity antibodies directed against conserved targets. Advanced antibody engineering techniques have extensive applications in the fields of immunology, biotechnology, diagnostics, and therapeutic medicines. However, there is limited knowledge regarding dynamic antibody development approaches. Therefore, this review extends beyond our understanding of conventional polyclonal and monoclonal antibodies. Furthermore, recent advances in antibody engineering techniques together with antibody fragments, display technologies, immunomodulation, and broad applications of antibodies are discussed to enhance innovative antibody production in pursuit of a healthier future for humans.

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Section: Single-chain Variable Fragment (Scfv)mentioning

confidence: 99%

Section: Library Construction Of Scfvmentioning

confidence: 99%

Antibody Engineering for Pursuing a Healthier Future

et al. 2017

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“…Given the occurrence of TTXs in European bivalve shellfish and the threat that this hazardous toxin poses to human health, the development of rapid, specific, sensitive, reliable and easyto-use methods for their detection is a matter of utmost importance. Accordingly, several methods have been reported for the detection of TTXs, including immunoassays [23][24][25][26][27][28][29][30][31][32][33] and immunosensors [23,[34][35][36][37][38][39][40][41]. Although some of these immunochemical approaches have been applied to the analysis of puffer fish [32,33,36,38,[40][41], newts [26], caddisflies [27], terrestrial flatworms [29], sea snails [38], urine [36] and milk/apple juice [37], none of them has been applied to the analysis of bivalve mollusks.…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels

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Rambla-Alegre

Leonardo

et al. 2018

Talanta

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The recent detection of tetrodotoxins (TTXs) in puffer fish and shellfish in Europe highlights the necessity to monitor the levels of TTXs in seafood by rapid, specific, sensitive and reliable methods in order to protect human consumers. A previous immunoassay for TTX detection in puffer fish, based on the use of self-assembled monolayers (SAMs) for the immobilization of TTX on maleimide plates (mELISA), has been modified and adapted to the analysis of oyster and mussel samples. Changing dithiol for cysteamine-based SAMs enabled reductions in the assay time and cost, while maintaining the sensitivity of the assay. The mELISA showed high selectivity for TTX since the antibody did not cross-react with co-occurring paralytic shellfish poisoning (PSP) toxins and no interferences were observed from arginine (Arg). Moreover, TTX-coated maleimide plates stored for 3 months at -20°C and 4°C were stable, thus when pre-prepared, the time to perform the assay is reduced. When analyzing shellfish samples, matrix effects and toxin recovery values strongly depended on the shellfish type and the sample treatment. Blank oyster extracts could be directly analyzed without solid-phase extraction (SPE) clean-up, whereas blank mussel extracts showed strong matrix effects and SPE and subsequent solvent evaporation were required for removal. However, the SPE clean-up and evaporation resulted in toxin loss. Toxin recovery values were taken as correction factors (CFs) and were applied to the quantification of TTX contents in the analysis of naturally-contaminated shellfish samples by mELISA. The lowest effective limits of detection (eLODs) were about 20 and 50µg/kg for oyster extracts without and with SPE clean-up, respectively, and about 30µg/kg for mussel extracts with both protocols, all of them substantially below the eLOD attained in the previous mELISA for puffer fish (230µg/kg). Analysis of naturally-contaminated samples by mELISA and comparison with LC-MS/MS quantifications demonstrated the viability of the approach. This mELISA is a selective and sensitive tool for the rapid detection of TTX in oyster and mussel samples showing promise to be implemented in routine monitoring programs to protect human health.

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“…Nevertheless, it is necessary to mention that, even though the detection was direct, a secondary Ab (non-labelled) was used to improve the anti-TTX MAb orientation and, therefore, the analysis time was still compromised. Other works have conjugated HRP [ 221 , 227 ] or ALP [ 223 , 228 ] to the anti-TTX antibody to avoid the use of a labelled secondary Ab. Despite the advantages of using conjugates, their stability (of the linkage and of the enzyme activity) still is a pending task.…”

Section: Biosensors Biochemical Assays and Cell-based Assaysmentioning

confidence: 99%

Alternative Methods for the Detection of Emerging Marine Toxins: Biosensors, Biochemical Assays and Cell-Based Assays

et al. 2014

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The emergence of marine toxins in water and seafood may have a considerable impact on public health. Although the tendency in Europe is to consolidate, when possible, official reference methods based on instrumental analysis, the development of alternative or complementary methods providing functional or toxicological information may provide advantages in terms of risk identification, but also low cost, simplicity, ease of use and high-throughput analysis. This article gives an overview of the immunoassays, cell-based assays, receptor-binding assays and biosensors that have been developed for the screening and quantification of emerging marine toxins: palytoxins, ciguatoxins, cyclic imines and tetrodotoxins. Their advantages and limitations are discussed, as well as their possible integration in research and monitoring programs.

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Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX

Cited by 36 publications

References 57 publications

Antibody Engineering for Pursuing a Healthier Future

Antibody Engineering for Pursuing a Healthier Future

Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels

Alternative Methods for the Detection of Emerging Marine Toxins: Biosensors, Biochemical Assays and Cell-Based Assays

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