Production of scFv recombinant fragments against 2,4-dichlorophenoxyacetic acid hapten using naďve phage library

Brichta, J.; Veselá, H; Fránek, Milan

doi:10.17221/5776-vetmed

Cited by 12 publications

(5 citation statements)

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“…Particularly, the production of RAbs to agrochemicals such as paraquat (Graham et al 1995), diuron (Scholthof et al 1997), parathion (Garret et al 1997), cyclohexanedione (Webb et al 1997), picloram (Yau et 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 al. 1998), chlorpyrifos (Alcocer et al 2000), atrazine (Charlton et al 2001), malathion (Nishi et al 2003), 2,4−dichlorophenoxyacetic acid (Brichta et al 2003), simetryn (Nishi et al 2005), and methamidophos (Li et al 2006) has been described. Initially, hapten-specific RAbs were obtained from hybridoma cells; subsequently, immunized lymphocytes were the source of genetic material to construct immune phage-display antibody libraries from which hapten-specific RAbs have been selected (Brichta et al 2005;Markus et al 2011).…”

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confidence: 99%

“…Initially, hapten-specific RAbs were obtained from hybridoma cells; subsequently, immunized lymphocytes were the source of genetic material to construct immune phage-display antibody libraries from which hapten-specific RAbs have been selected (Brichta et al 2005;Markus et al 2011). Lately, hapten-specific RAbs have also been isolated from non-immune or even synthetic libraries (Brichta et al 2003;Kramer and Hock 2007;Markus et al 2011). The most usual recombinant antibody fragment was the single chain variable fragment (scFv), where the V H and V L chains are single-domain linked by a 15 amino acids long polypeptide linker (Brichta et al 2005).…”

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Comparative Study of Monoclonal and Recombinant Antibody-Based Immunoassays for Fungicide Analysis in Fruit Juices

et al. 2013

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Abstract:A comparative study of the analytical performance of enzyme-linked immunosorbent assays (ELISAs), based on monoclonal and recombinant antibodies, for the determination of fungicide residues in fruit juices has been carried out. To this aim, three murine hybridoma cell lines secreting specific monoclonal antibodies against tetraconazole, thiabendazole, and imazalil were used as a source of immunoglobulin gene fragments for the production of single-chain variable fragment (scFv) and fusion scFv-pIII recombinant antibodies in Escherichia coli. Selected recombinant antibodies displayed cross-reactivity profiles very similar to those of the parent monoclonal antibodies. Imazalil and tetraconazole recombinant antibodies showed one order of magnitude lower affinity than their respective monoclonal antibodies, whereas the thiabendazole recombinant antibodies showed an affinity similar to that of their parent monoclonal antibody. On the other hand, scFv-pIII fusion fragments showed similar analytical properties as, and occasionally better than, scFv recombinant antibodies. Finally, ELISAs developed from each antibody type showed similar analytical performance when applied to the analysis of the target fungicides in fruit juices. Powered by Editorial Manager® and Preprint Manager® from Aries Systems Corporation AbstractA comparative study of the analytical performance of enzyme-linked immunosorbent assays (ELISAs), based on monoclonal and recombinant antibodies, for the determination of fungicide residues in fruit juices has been carried out. To this aim, three murine hybridoma cell lines secreting specific monoclonal antibodies against tetraconazole, thiabendazole, and imazalil were used as a source of immunoglobulin gene fragments for the production of single-chain variable fragment (scFv) and fusion scFv-pIII recombinant antibodies in Escherichia coli. Selected recombinant antibodies displayed cross-reactivity profiles very similar to those of the parent monoclonal antibodies. Imazalil and tetraconazole recombinant antibodies showed one order of magnitude lower affinity than their respective monoclonal antibodies, whereas the thiabendazole recombinant antibodies showed an affinity similar to that of their parent monoclonal antibody. On the other hand, scFv-pIII fusion fragments showed similar analytical properties as, and occasionally better than, scFv recombinant antibodies. Finally, ELISAs developed from each antibody type showed similar analytical performance when applied to the analysis of the target fungicides in fruit juices.

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Comparative Study of Monoclonal and Recombinant Antibody-Based Immunoassays for Fungicide Analysis in Fruit Juices

et al. 2013

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“…Mutations hampering production and/or leading to aggregation and inclusion body formation have been eliminated. Optimal numbers of selection cycles and use of free haptens as elution agents have been reported to improve the library selection output Strachan et al, 2002;Brichta et al, 2003). Moreover the efficiency of selection process can be largely increased by using selectively infecting phage particles.…”

Section: Resultsmentioning

confidence: 99%

“…The Griffin1 library was also used for preparation of phage antibodies against herbicide 2,4-dichlorophenoxyacetic acid. Brichta et al (2003) panned the Griffin library against 2,4-D herbicide. Four cycles of panning were used in the selection process and free 2,4-D hapten molecules was used during the elution step.…”

Section: Single-pot Librariesmentioning

confidence: 99%

Generation of hapten-specific recombinant antibodies: antibody phage display technology: a review

Brichta¹,

Hnilova²,

Viskovic³

2005

Vet. Med.

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Production of antibodies has been revolutionized by the development of modern molecular biology methods for the expression of recombinant DNA. Phage display technology represents one of the most powerful tools for production and selection of recombinant antibodies and has been recognized as a valuable alternative way for the preparation of antibodies of a desired specificity. In comparison to poly-and monoclonal antibodies, recombinant antibodies using the phage display technology can be prepared faster, in more automatic process and with reduced consumption of laboratory animals. This review summarizes current trends of phage display technology with focus on the generation of hapten-specific recombinant antibodies and gives the examples of successful applications of phage display in the environmental analysis of low molecular weight compound.

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“…Purified NUsc1 was stored in phosphate-buffered saline (PBS) (Dulbecco's PBS without calcium or magnesium; Corning), pH 7.4 at À80°C. After analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and SEC (size-exclusion chromatography), the following were added to NUsc1 for cryoprotection: 15% glycerol, 0.1% casein, and 1 mM PMSF (phenylmethane sulfonyl fluoride) (Brichta et al 2003).…”

Section: Production Of Phage-free Nusc1mentioning

confidence: 99%

A human scFv antibody that targets and neutralizes high molecular weight pathogenic amyloid‐β oligomers

Sebollela

Cline

Popova

et al. 2017

Journal of Neurochemistry

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Brain accumulation of soluble oligomers of the amyloid-β peptide (AβOs) is increasingly considered a key early event in the pathogenesis of Alzheimer's disease (AD). A variety of AβO species have been identified, both in vitro and in vivo, ranging from dimers to 24mers and higher order oligomers. However, there is no consensus in the literature regarding which AβO species are most germane to AD pathogenesis. Antibodies capable of specifically recognizing defined subpopulations of AβOs would be a valuable asset in the identification, isolation, and characterization of AD-relevant AβO species. Here, we report the characterization of a human single chain antibody fragment (scFv) denoted NUsc1, one of a number of scFvs we have identified that stringently distinguish AβOs from both monomeric and fibrillar Aβ. NUsc1 readily detected AβOs previously bound to dendrites in cultured hippocampal neurons. In addition, NUsc1 blocked AβO binding and reduced AβO-induced neuronal oxidative stress and tau hyperphosphorylation in cultured neurons. NUsc1 further distinguished brain extracts from AD-transgenic mice from wild type (WT) mice, and detected endogenous AβOs in fixed AD brain tissue and AD brain extracts. Biochemical analyses indicated that NUsc1 targets a subpopulation of AβOs with apparent molecular mass greater than 50 kDa. Results indicate that NUsc1 targets a particular AβO species relevant to AD pathogenesis, and suggest that NUsc1 may constitute an effective tool for AD diagnostics and therapeutics.

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Production of scFv recombinant fragments against 2,4-dichlorophenoxyacetic acid hapten using naďve phage library

Cited by 12 publications

References 28 publications

Comparative Study of Monoclonal and Recombinant Antibody-Based Immunoassays for Fungicide Analysis in Fruit Juices

Comparative Study of Monoclonal and Recombinant Antibody-Based Immunoassays for Fungicide Analysis in Fruit Juices

Generation of hapten-specific recombinant antibodies: antibody phage display technology: a review

A human scFv antibody that targets and neutralizes high molecular weight pathogenic amyloid‐β oligomers

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