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首页> 外文期刊>Developmental biology >Monoclonal antibody to human fibronectin: Production and characterization using human muscle cultures☆
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Monoclonal antibody to human fibronectin: Production and characterization using human muscle cultures☆

机译:人纤连蛋白单克隆抗体:利用人的肌肉培养物进行生产和鉴定☆

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Amonoclonalanti-humanfibronectinantibody(2.3F9)wasproducedfromacellfusioninvolvingthespleencellsfromaBALBcmouseimmunizedwithhumanmyoblastcellsandP3X63Ag8mousemyelomacells.2.3F9antibodyisoftheIgMclass.Radioimmunoassayshowedthat2.3F9antibodyboundtohumanskinfibroblastsandmusclecellsbutnottoperipherallymphocytes,redbloodcells,orthelymphoblastoidcelllineBristol8,showingthattheantigenisnotpresentonallhumancells.Ouchterlonyimmunodiffusionanalysisrevealedaprecipitinlinewithplasmafibronectin(coldinsolubleglobulin).Immunoprecipitationwithantibody2.3F9fromNP40extractsof[35S]methioninehumanmuscleculturesrevealedthespecificprecipitationofaproteinof220,000molecularweightthatcomigratedwithfibronectin.Theantibodyisnotspeciesspecificandculturesofratmusclecellswerestainedusingindirectimmunofluorescence,althoughthestainingwasweak.Thecross-reactivitymaythereforenotbecompleteacrossspecies.Humanmyoblastsarelowinfibronectinbutastheculturesdivideandformmyotubesthelevelincreasesdramatically.Themyotubesaredevoidoffibronectin,however,andthemajorityofstainingisassociatedwiththemononucleatedcells.Theseresultswereconfirmedusingamyogenichumanmuscleclonewheremyoblastswerefoundtosynthesizeandaccumulatefibronectinwhilemyotubesdidnot.Immunofluorescentstainingoftransverseandlongitudinalsectionsofhumanmuscledetectedfibronectinontheepimysium,endomysium,andperimysium.
机译:Amonoclonalanti-humanfibronectinantibody(2.3F9)wasproducedfromacellfusioninvolvingthespleencellsfromaBALBcmouseimmunizedwithhumanmyoblastcellsandP3X63Ag8mousemyelomacells.2.3F9antibodyisoftheIgMclass.Radioimmunoassayshowedthat2.3F9antibodyboundtohumanskinfibroblastsandmusclecellsbutnottoperipherallymphocytes,redbloodcells,orthelymphoblastoidcelllineBristol8,showingthattheantigenisnotpresentonallhumancells.Ouchterlonyimmunodiffusionanalysisrevealedaprecipitinlinewithplasmafibronectin(coldinsolubleglobulin).Immunoprecipitationwithantibody2.3F9fromNP40extractsof [35S] methioninehumanmuscleculturesrevealedthespecificprecipitationofaproteinof220,000molecularweightthatcomigratedwithfibronectin.Theantibodyisnotspeciesspecificandculturesofratmusclecellswerestainedusingindirectimmunofluorescence,althoughthestainingwasweak.Thecross-reactivitymaythereforenotbecompleteacrossspecies.Humanmyoblastsarelowinfibronectinbutastheculturesdivideandformmyotubesthelevelincreasesdramatically.Themyotubesaredevoi但是,多巴尼单抗和多数染色与单核细胞有关。通过使用肌原性人肌克隆证实了上述结果,其中成肌细胞在整个肌管中被合成,并在横断面和长的免疫荧光上没有被检测到。

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