The change of the scFv into the Fab format improves the stability and in vivo toxin neutralization capacity of recombinant antibodies

Quintero-Hernández, Verónica; Juárez-González, Víctor Rivelino; Ortíz-León, Mauricio; Sánchez, Rosalba; Possani, Lourival D.; Becerril, Baltazar

doi:10.1016/j.molimm.2006.05.009

Cited by 77 publications

(36 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, they also have the disadvantages of high cost, varying stability, and challenging production and purification (Worn and Pluckthun 2001, Quintero-Hernandez et al 2007, Malpiedi et al 2013. Antibody fragments obtained by chemical reduction are an  2-MEA -2-mercaptoethylamine EDTA -ethylenediaminetetraacetic acid disodium dihydrate TCEP -(2-carboxyethyl)phosphine attractive alternative as their production is inexpensive and uncomplicated.…”

Section: Introductionmentioning

confidence: 99%

Considerations in producing preferentially reduced half-antibody fragments

Makaraviciute

Jackson

Millner

et al. 2016

Journal of Immunological Methods

View full text Add to dashboard Cite

Half-antibody fragments are a promising reagent for biosensing, drug-delivery and labeling applications, since exposure of the free thiol group in the Fc hinge region allows oriented Preferential reduction of rabbit anti-myoglobin IgG antibodies was optimized and the highest half-antibody yield was obtained with 35 mM TCEP. Finally, it has been demonstrated that produced anti-myoglobin half-IgG fragments retained their binding activity.

show abstract

Section: Introductionmentioning

confidence: 99%

Considerations in producing preferentially reduced half-antibody fragments

Makaraviciute

Jackson

Millner

et al. 2016

Journal of Immunological Methods

View full text Add to dashboard Cite

show abstract

“…Finally, antisera to be used as antidotes (antivenoms) by humans need to be highly potent and specific to trap the rapidly diffusing small toxins and still be devoid of potential secondary effects. Hence, and although some studies pointed to the higher functional stability and neutralizing capacity of antigen binding fragment (Fab) 5 molecules compared with their single-chain variable-fragment antibody (scFv) counterparts (2,3), most strategies were aimed at generating, by protein and/or peptide engineering, new molecules such as recombinant Fab, scFv, tandem scFv, diabody or nanobody molecules, with enhanced recognition properties (Refs. 4 -15; for review, see Refs.…”

mentioning

confidence: 99%

Structural Insights into Antibody Sequestering and Neutralizing of Na+ Channel α-Type Modulator from Old World Scorpion Venom

Fabrichny

Mondielli

Conrod

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background:The antigenic specificity of scorpion ␣-toxins, which target Na v channels, hinders efficient antiserum production. Results: Structures of two antibodies, which together neutralize the main toxins in a threatening venom, were solved in toxin-bound and unbound forms, respectively. Conclusion: Selective toxin trapping involves distinctive molecular determinants and bound toxin orientations. Significance: These complementary templates will help design new neutralizing molecules suitable for immunotherapy.

show abstract

“…Adding mammalian antibody domains has been used to alter the characteristics of scFvs from various sources [36][37][38]. The present study has shown that the vectors and approaches developed by Greunke et al [24] to create larger bivalent molecules from monovalent chicken antibody fragments can be used for improving avian scFvs which under-perform in certain immunoassay applications.…”

Section: Discussionmentioning

confidence: 76%

Chicken scFvs and bivalent scFv-CH fusions directed against HSP65 of Mycobacterium bovis

et al. 2010

View full text Add to dashboard Cite

a b s t r a c tTwo chicken single-chain variable region antibody fragments (scFvs) that recognised the 65 kDa heatshock protein (HSP65) of Mycobacterium bovis were selected from a large semi-synthetic phage displayed library. Both recognised HSP65 in indirect enzyme-linked immunosorbent assay (ELISA) and immunoblots and retained their activity during storage. Neither, however, could function as the capture reagent in a sandwich ELISA when immobilised on polystyrene. To establish whether they could be engineered for general use in immunotests, the genes coding for these scFvs were subcloned in expression vectors that contained sequences encoding chicken IgY heavy-chain constant region domains. This resulted in larger bivalent constructs which more closely resembled IgY molecules. The engineered fragments were evaluated in ELISAs and gold-conjugated immunochromatographic tests (ICTs). In contrast to their previous behaviour as scFvs, the modified fragments (designated ''gallibodies'') could be used for immunocapture in ELISA and could be readily conjugated to colloidal gold nanoparticles. A sandwich ICT that could detect recombinant HSP65 was also devised. Although converting the recombinant single-chain monomeric antibody fragments to bivalent immunoglobulin-like molecules did not entirely 'standardise' the behaviour of the scFvs, this approach remains potentially useful for developing practical, robust, immunodiagnostic reagents.

show abstract

The change of the scFv into the Fab format improves the stability and in vivo toxin neutralization capacity of recombinant antibodies

Cited by 77 publications

References 24 publications

Considerations in producing preferentially reduced half-antibody fragments

Considerations in producing preferentially reduced half-antibody fragments

Structural Insights into Antibody Sequestering and Neutralizing of Na+ Channel α-Type Modulator from Old World Scorpion Venom

Chicken scFvs and bivalent scFv-CH fusions directed against HSP65 of Mycobacterium bovis

Contact Info

Product

Resources

About