Recombinant antibodies and their use for food immunoanalysis

Peltomaa, Riikka; Barderas, Rodrigo; Benito‐Peña, Elena; Moreno‐Bondi, María C.

doi:10.1007/s00216-021-03619-7

Cited by 34 publications

(21 citation statements)

References 148 publications

(179 reference statements)

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“…Single-chain fragment variables (scFvs) have several advantages over antibodies, including their small size and their ease of production in bacteria (Peltomaa et al, 2022; Weisser and Hall, 2009). We generated a His-tagged scFv based on the sequence of an antibody targeting DLL3 (Methods) ( Figure 2A and Supplementary Figure S2A ).…”

Section: Resultsmentioning

confidence: 99%

Dll3 Regulates Notch Signaling in SCLC

Kim

Sage

2022

Preprint

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Tumor heterogeneity plays a critical role in tumor development and response to treatment. In small-cell lung cancer (SCLC), intratumoral heterogeneity is driven in part by the Notch signaling pathway, which reprograms neuroendocrine cancer cells to a less/non-neuroendocrine state. Here we investigated the atypical Notch ligand, DLL3 as a biomarker of the neuroendocrine state and a regulator of cell-cell interactions in SCLC. We first built a mathematical model to predict the impact of DLL3 expression on SCLC cell populations. We next tested this model using a single-chain variable fragment (scFv) to track DLL3 expression in vivo and a new mouse model with inducible expression of DLL3 in SCLC tumors. We found that high levels of DLL3 promote the expansion of a new SCLC cell population with lower expression levels of both neuroendocrine and non-neuroendocrine markers. This work may influence how DLL3-targeting therapies are used in SCLC patients.

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Section: Resultsmentioning

confidence: 99%

Dll3 Regulates Notch Signaling in SCLC

Kim

Sage

2022

Preprint

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“…In general, scFv exhibits various advantages, including rapid and inexpensive animal-free production, high potential for improving binding properties, and easy transformation into multispecific molecules to achieve multiple biological effects. Thus, a number of scientists have proposed that scFv can replace traditional animal-derived antibodies; however, others believe that the preparation and application technology of scFv is not mature enough, F I G U R E 1 Basic structure of typical immunoglobulin G (IgG) antibody and scFv antibody and scFv should be used as a supplementary immune reagent, and not as a substitute (Peltomaa et al, 2022). Currently, scFvs are widely used in clinical treatment; however, their application in the field of food safety must be strengthened (Lei et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

“…Currently, the proficient application of phage display technology with the unique advantages of high throughput, high efficiency, short development cycle, and low cost has promoted the single‐chain fragment variable (scFv) to become one of the most widely used recombinant antibody formats (Roth et al., 2021). Compared with other types of recombinant antibodies, scFv maintains the diversity of heavy and light chain binding, high presentation efficiency on the phage surface, and easy expression in most expression systems (Peltomaa et al., 2022). A typical antibody consists of two light chains (∼24 kDa) and two heavy chains (∼55 kDa) linked together by disulfide and noncovalent bonds (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

Single‐chain fragment variable produced by phage display technology: Construction, selection, mutation, expression, and recent applications in food safety

et al. 2022

Comp Rev Food Sci Food Safe

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Immunoassays are reliable, efficient, and accurate methods for the analysis of small‐molecule harmful substances (such as pesticides, veterinary drugs, and biological toxins) that may be present in food. However, traditional polyclonal and monoclonal antibodies are limited by animal hosts and hinder further development of immunoassays. With the gradual application of phage display technology as an efficient in vitro selection technology, the single‐chain fragment variable (scFv) now provides an exciting alternative to traditional antibodies. Efficiently constructed scFv source libraries and specifically designed biopanning schemes can now yield scFvs possessing specific recognition capabilities. A rational mutation strategy further enhances the affinity of scFv, and allows it to reach a level that cannot be achieved by immunization. Finally, appropriate prokaryotic expression measures ensure stable and efficient production of scFv. Therefore, when developing excellent scFvs, it is necessary to focus on three key aspects of this process that include screening, mutation, and expression. In this review, we analyze in detail the preparation and affinity improvement process for scFv and provide insights into the research progress and development trend of scFv‐based immunoassay methods for monitoring small‐molecule harmful substances.

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“…With the rapid development of molecular biology research for antibodies, increasingly more genetically engineered antibodies to detect small molecule compounds have been developed [ 18 ]. Recombinant antibodies such as fragment of antigen binding (Fab) and single-chain antibody fragment (ScFv) have been utilized in ELISAs and biosensors [ 19 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

A Nanobody-Based Immunoassay for Detection of Ustilaginoidins in Rice Samples

Wang

Yin

et al. 2022

Toxins

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Ustilaginoidins are a class of bis-naphtho-γ-pyrone mycotoxins produced by the pathogen Villosiclava virens of rice false smut, which has recently become one of the most devastating diseases in rice-growing regions worldwide. In this research, the nanobody phage display library was established after an alpaca was immunized with the hemiustilaginoidin F-hapten coupled with bovine serum albumin (BSA). Heterologous antigen selection and combing trypsin with competition alternant elution methods were performed for nanobody screening. Two nanobodies, namely, Nb-B15 and Nb–C21, were selected for the establishment of indirect competitive enzyme-linked immunosorbent assays (ic-ELISAs). For Nb–B15 and Nb-C21, their IC50 values were 11.86 μg/mL and 11.22 μg/mL, and the detection ranges were at 3.41–19.98 μg/mL and 1.17–32.13 μg/mL, respectively. Two nanobodies had a broad spectrum to quantify the contents of total ustilaginoidins in rice samples according to cross-reactivity. The recognition mechanisms of Nb-B15 and Nb-C21 against ustilaginoidin A were elucidated by molecular modeling and docking. The key amino acid sites for the binding of Nb–B15 or Nb–C21 to ustilaginoidin A were mainly located in the FR1 and CDR1 regions. As Nb-B15 was superior to Nb–C21 in the aspects of protein expression, ELISA titer, and tolerance to organic solvents, it was selected for application in the detection of actual contaminated rice samples. The total ustilaginoidin contents of rice samples were analyzed by Nb–B15-based ic–ELISA and HPLC-DAD, between which the results were found to be consistent. The developed immunoassay based on the nanobody from the alpaca can be employed as a rapid and effective method for detection of total utilaginoidins in contaminated rice samples.

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Recombinant antibodies and their use for food immunoanalysis

Cited by 34 publications

References 148 publications

Dll3 Regulates Notch Signaling in SCLC

Dll3 Regulates Notch Signaling in SCLC

Single‐chain fragment variable produced by phage display technology: Construction, selection, mutation, expression, and recent applications in food safety

A Nanobody-Based Immunoassay for Detection of Ustilaginoidins in Rice Samples

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