Single-Chain Fv of Anti-idiotype 11-1G10 Antibody Interacts with Antibody NC41 Single-Chain Fv with a Higher Affinity than the Affinity for the Interaction of the Parent Fab Fragments

Iliades, Peter; Dougan, David A.; Oddie, Geoffrey W.; Metzger, Dennis W.; Hudson, Peter J.; Kortt, Alexander A.

doi:10.1023/a:1022584702108

Cited by 16 publications

(2 citation statements)

References 2 publications

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“…The V H and V L genes were amplified by PCR from the parent NC10 [13]and 11‐1G10 [14]hybridomas and scFvs were constructed by PCR overlap extension. NC10 scFv‐5 was constructed using a Gly 4 Ser five‐residue linker [8]and the scFv‐0 genes were constructed by ligation between codons for C‐terminal V H ‐Ser 112 and N‐terminal V L ‐Asp 1 for NC10 and between C‐terminal V H ‐Ser 113 and N‐terminal V L ‐Gln 1 for 11‐1G10.…”

Section: Methodsmentioning

confidence: 99%

Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments

Lawrence¹,

Kortt

Iliades

et al. 1998

FEBS Letters

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Electron microscopy of dimeric and trimeric single chain antibody Fv fragments (scFvs) complexed with antiidiotype Fab fragments was used to reveal the orientation of antigen binding sites. This is the first structural analysis that discloses the multivalent binding orientation of scFv trimers (triabodies). Three different scFv molecules were used for the imaging analysis; NC10 scFv-5 and scFv-0, with five-and zeroresidue linkers respectively between the V r and V v domains, were complexed with 3-2G12 anti-idiotype Fab fragments and 11-1G10 scFv-0 was complexed with NC41 anti-idiotype Fab fragments. The scFv-5 molecules formed bivalent dimers (diabodies) and the zero-linker scFv-0 molecules formed trivalent trimers (triabodies). The images of the NC10 diabody-Fab complex appear as boomerangs, not as a linear molecule, with a variable angle between the two Fab arms and the triabody-Fab complexes appear as tripods.z 1998 Federation of European Biochemical Societies.

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Section: Methodsmentioning

confidence: 99%

Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments

Lawrence¹,

Kortt

Iliades

et al. 1998

FEBS Letters

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“…This is surprising as both fragments bind to identical residues in the gp120 binding site. Only in few cases ScFvs have shown to bind with elevated affinity than the linked Fab [21] . The plausible reasons might be the multimerization that enhances the affinity for solube Scfabs.…”

Section: Discussionmentioning

confidence: 99%

Surface plasmon resonance (SPR) based binding studies of refolded single chain antibody fragments

Singh

2018

Biochemistry and Biophysics Reports

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Recent advances in Recombinant antibody technology / Antibody Engineering has given impetus to the genetic manipulation of antibody fragments that has paved the way for better understanding of the structure and functions of immunoglobulins and also has escalated their use in immunotherapy. Bacterial expression system such as Escherichia coli has complemented this technique through the expression of recombinant antibodies. Present communication has attempted to optimize the expression and refolding protocol of single chain fragment variable (ScFv) and single chain antigen binding fragment (ScFab) using E.coli expression system. Efficiency of refolding protocol was validated by structural analysis by CD, native folding by fluorescence and functional analysis by its binding with full length HIV-1 gp120 via SPR. Results show the predominant β–sheet (CD) as secondary structural content and native folding via red shift (tryptophan fluorescence). The single chain fragments have shown good binding with HIV-1 gp120 thus validating the expression and refolding strategy and also reinstating E.coli as model expression system for recombinant antibody engineering. SPR based binding analysis coupled with E.coli based expression and purification will have implication for HIV therapeutics and will set a benchmark for future studies of similar kind.

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Survey of the 1998 optical biosensor literature

Myszka

1999

J. Mol. Recognit.

120

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The utilization of optical biosensors to study molecular interactions continues to expand. In 1998, 384 articles relating to the use of commercial biosensors were published in 130 different journals. While significant strides in new applications and methodology were made, a majority of the biosensor literature is of rather poor quality. Basic information about experimental conditions is often not presented and many publications fail to display the experimental data, bringing into question the credibility of the results. This review provides suggestions on how to collect, analyze and report biosensor data.

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Single-Chain Fv of Anti-idiotype 11-1G10 Antibody Interacts with Antibody NC41 Single-Chain Fv with a Higher Affinity than the Affinity for the Interaction of the Parent Fab Fragments

Cited by 16 publications

References 2 publications

Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments

Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments

Surface plasmon resonance (SPR) based binding studies of refolded single chain antibody fragments

Survey of the 1998 optical biosensor literature

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