Stable expression of cloned human antibody genes in murine myeloma cells

Knight, David; McDonough, Margaret; Moore, Maria Arevalo; Abercrombie, David M.; Siegel, Richard D.; Ghrayeb, John

doi:10.3233/hab-1992-3303

Cited by 10 publications

(5 citation statements)

References 29 publications

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“…Human monoclonal antibodies (mAbs) are a new class of diagnostic and therapeutic agents which have made a substantial impact in the biomedical field. They are less likely to provoke the immune response which can severely impair the clinical efficiency of murine antibodies [1]. Indeed, human anti‐mouse antibodies (HAMA) may accelerate the clearance of the injected mAb, block its binding to the antigen and/or trigger allergic reactions.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the V genes coding for a monospecific human antibody to myosin and functional expression of single chain Fv fragments

Laroche‐Traineau

Jacobin²,

Biard-Piechaczyk

et al. 1999

FEBS Letters

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A monospecific human IgM monoclonal antibody (mAb), reactive with myosin from human heart, has been obtained by EBV transformation. This mAb may have a diagnostic potential in the imaging of myocardial necrosis. However, owing to the fact that the molecular mass of an IgM is 900 kDa, a poor diffusion and a slow penetration inside necrotic myocytes could reduce its capacity for scintigraphic detection. In order to alleviate these problems, we constructed the scFv by cloning the VH and VL domains into the pHOG21 vector. Analysis of the V genes proved an unmutated configuration showing that the immortalized B cell issued from the primary IgM repertoire. The expression product in Escherichia coli was a 35 kDa scFv fragment with the antigen-binding specificity of the parental mAb.z 1999 Federation of European Biochemical Societies.

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Section: Introductionmentioning

confidence: 99%

Analysis of the V genes coding for a monospecific human antibody to myosin and functional expression of single chain Fv fragments

Laroche‐Traineau

Jacobin²,

Biard-Piechaczyk

et al. 1999

FEBS Letters

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“…Traditional methods for obtaining HmAbs (i.e., immortalization of B cells by cell fusion or transformation) can result in a low and unstable Ab secretion. 7 Several new methods have been devised for the direct cloning of human variable region genes via polymerase chain reaction and phage combinatorial libraries. 8,9 Both types of HmAb production can benefit from expression systems that support the stable high-level Ab secretion required for therapeutic use.…”

mentioning

confidence: 99%

Generation, immunologic characterization and antitumor effects of human monoclonal antibodies for carcinoembryonic antigen

Imakiire

Shibaguchi

Abe

et al. 2003

Intl Journal of Cancer

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“…To establish further hapten‐specific antibody formats suitable for standard procedures, we aimed for construction of both scFv‐based IgY constructs as well as chimaeric IgY, which allowed a reliable evaluation in standard assays for quantification of TNT. Several studies have demonstrated that the conversion of antibody fragments and the expression of resulting mammalian immunoglobulins in mammalian cell lines of myeloid or non‐myeloid origin result in fully functional immunoglobulins [34–36]. Consequently, transfection of the human cell line HEK‐293 resulted in stable secretion of the majority of the IgY antibodies.…”

Section: Discussionmentioning

confidence: 99%

Establishment of hapten‐specific monoclonal avian IgY by conversion of antibody fragments obtained from combinatorial libraries

Deckers

Braren

Greunke

et al. 2009

Biotech and App Biochem

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Nowadays, recombinant antibody and phage display technology enable the efficient generation of immunotools and a subsequent manipulation for optimized affinity, specificity or overall performance. Such advantages are of particular interest for haptenic target structures, such as TNT (2,4,6-trinitrotoluene). The toxicity of TNT and its breakdown products makes a reliable and fast detection of low levels in aqueous samples highly important. In the present study, we aimed for the generation of scFvs (single-chain antibody fragments) specific for the TNT-surrogate TNP (2,4,6-trinitrophenyl) and their subsequent production as monoclonal avian IgY immunoglobulins providing improved assay performance. Therefore we subjected a human synthetic scFv library to selection following different strategies. TNP-specific human antibody fragments could be identified, characterized for their primary structure and evaluated for production as soluble scFv in Escherichia coli. Additionally, a murine TNP-specific antibody fragment was obtained from the hybridoma 11B3; however, the prokaryotic expression level was found to be limited. To generate and evaluate immunoglobulin formats with superior characteristics, all recombinant antibody fragments then were converted into two different chimaeric bivalent IgY antibody formats. After expression in mammalian cells, the IgY antibodies were assessed for their reactivity towards TNT. The IgY antibodies generated on the basis of the combinatorial library proved to be useful for detection of TNT, thereby emphasizing the high potential of this approach for the development of detection devices for immunoassay-based techniques.

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Stable expression of cloned human antibody genes in murine myeloma cells

Cited by 10 publications

References 29 publications

Analysis of the V genes coding for a monospecific human antibody to myosin and functional expression of single chain Fv fragments

Analysis of the V genes coding for a monospecific human antibody to myosin and functional expression of single chain Fv fragments

Generation, immunologic characterization and antitumor effects of human monoclonal antibodies for carcinoembryonic antigen

Establishment of hapten‐specific monoclonal avian IgY by conversion of antibody fragments obtained from combinatorial libraries

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