Pub Date : 2001-02-01DOI: 10.1089/027245701300060481
Shu-Fen Chou, Chien-Yuan Chen
The aim of this study was to produce monoclonal and polyclonal antibodies against a nonspecific tumor marker, human ferritin. Hyperimmune ICR mice produced polyclonal antibodies after injection with 0.5 mL pristane, and were injected with NS-1 myeloma cells 2 weeks later. Hyperimmune Balb/c mice were used for the production of monoclonal antibodies (MAbs). Mice were immunized four times, given a final boost, and their spleen cells were collected and fused with NS-1 myeloma cells under the presence of PEG 1500. The fused cells were then selected in the HAT-RPMIX medium. Anti-ferritin antibody-secreting hybridoma cell lines with high titer were cloned by enzyme-linked immunoadsorbent assay (ELISA) and then subcloned by limiting dilution in 15% fetal bovine serum (FBS) HT-RPMIX medium. Five murine hybridoma-producing antiferritin MAbs were obtained and designated 1AD11F9, 1AD11E11, 2AD11D2, 2AD11A5, and 3AD11G8. Isotypes of these MAbs were identified as IgM heavy chain and kappa light chain. Hitrap Protein A and Hitrap IgM purification column were used for the purification of polyclonal and monoclonal antibodies, respectively.
本研究的目的是制备针对非特异性肿瘤标志物人铁蛋白的单克隆和多克隆抗体。高免疫ICR小鼠注射0.5 mL普里斯坦后产生多克隆抗体，2周后注射NS-1骨髓瘤细胞。采用高免疫Balb/c小鼠制备单克隆抗体(mab)。小鼠免疫四次，最后一次增强，收集脾脏细胞，在PEG 1500存在下与NS-1骨髓瘤细胞融合。然后在HAT-RPMIX培养基中选择融合细胞。采用酶联免疫吸附法(ELISA)克隆高效价的抗铁蛋白抗体分泌杂交瘤细胞系，然后在15%胎牛血清(FBS) HT-RPMIX培养基中限制性稀释亚克隆。获得了5种产生杂交瘤的抗铁蛋白单抗，分别命名为1AD11F9、1AD11E11、2AD11D2、2AD11A5和3AD11G8。这些单克隆抗体的同型分别为IgM重链和kappa轻链。采用Hitrap Protein A和Hitrap IgM纯化柱分别纯化多克隆抗体和单克隆抗体。
{"title":"Monoclonal and polyclonal antibodies against human ferritin, a nonspecific tumor marker.","authors":"Shu-Fen Chou, Chien-Yuan Chen","doi":"10.1089/027245701300060481","DOIUrl":"https://doi.org/10.1089/027245701300060481","url":null,"abstract":"The aim of this study was to produce monoclonal and polyclonal antibodies against a nonspecific tumor marker, human ferritin. Hyperimmune ICR mice produced polyclonal antibodies after injection with 0.5 mL pristane, and were injected with NS-1 myeloma cells 2 weeks later. Hyperimmune Balb/c mice were used for the production of monoclonal antibodies (MAbs). Mice were immunized four times, given a final boost, and their spleen cells were collected and fused with NS-1 myeloma cells under the presence of PEG 1500. The fused cells were then selected in the HAT-RPMIX medium. Anti-ferritin antibody-secreting hybridoma cell lines with high titer were cloned by enzyme-linked immunoadsorbent assay (ELISA) and then subcloned by limiting dilution in 15% fetal bovine serum (FBS) HT-RPMIX medium. Five murine hybridoma-producing antiferritin MAbs were obtained and designated 1AD11F9, 1AD11E11, 2AD11D2, 2AD11A5, and 3AD11G8. Isotypes of these MAbs were identified as IgM heavy chain and kappa light chain. Hitrap Protein A and Hitrap IgM purification column were used for the purification of polyclonal and monoclonal antibodies, respectively.","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 1 1","pages":"59-62"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701300060481","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60498611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-02-01DOI: 10.1089/027245701300060445
X. Tang, X. Zhang, H. Xu
A new treble-coated enzyme-linked immunoadsorbent assay (ELISA) kit of detecting Hepatitis B virus (HBV) surface antigen subtypes a, d and r (HBsAg-a, -d, -r) was developed by using four established hybridoma cell lines, of which two specifically secrete monoclonal antibodies (MAbs) against HBsAg-a (anti-HBsAg-a), one against -d (anti-HBsAg-d), and one against -r (anti-HBsAg-r). The approach of hybridoma cell lines' establishment were by fusing myeloma cells (SP2/0) with splenocytes from BALB/c mice immunized with a mixture of HBsAg-a, -d, -r. The ascitic MAb productivity of the four cell lines was at the titres of 1:10(6)-1:10(8). A treble-coated ELISA based HBV diagnostic kit was developed for detecting all of the three responding subtypes of HBsAgs. A 96-well ELISA microplate was coated with anti-HBSAg-a, -d, -r at a ratio of 3: 1: 0.5, with a horseradish peroxidase (HRP) conjugated anti-HBsAg-a as the labelled antibody. For clinical application, the new developed diagnostic kit detected HBsAgs of adr, adw, ayr, and ayw at a rate of lower than 0.25, 0.25, 0.5, and 0.5 ng/mL, respectively. Results indicated that this kit was more rapid and sensitive than that other current ELISA-based kits coated with a single MAb (e.g., anti-HBsAg-a).
{"title":"Establishment of hybridoma cell lines producing monoclonal antibodies against hepatitis B virus surface antigens (a, d, and r) and development of sensitive ELISA diagnostic test.","authors":"X. Tang, X. Zhang, H. Xu","doi":"10.1089/027245701300060445","DOIUrl":"https://doi.org/10.1089/027245701300060445","url":null,"abstract":"A new treble-coated enzyme-linked immunoadsorbent assay (ELISA) kit of detecting Hepatitis B virus (HBV) surface antigen subtypes a, d and r (HBsAg-a, -d, -r) was developed by using four established hybridoma cell lines, of which two specifically secrete monoclonal antibodies (MAbs) against HBsAg-a (anti-HBsAg-a), one against -d (anti-HBsAg-d), and one against -r (anti-HBsAg-r). The approach of hybridoma cell lines' establishment were by fusing myeloma cells (SP2/0) with splenocytes from BALB/c mice immunized with a mixture of HBsAg-a, -d, -r. The ascitic MAb productivity of the four cell lines was at the titres of 1:10(6)-1:10(8). A treble-coated ELISA based HBV diagnostic kit was developed for detecting all of the three responding subtypes of HBsAgs. A 96-well ELISA microplate was coated with anti-HBSAg-a, -d, -r at a ratio of 3: 1: 0.5, with a horseradish peroxidase (HRP) conjugated anti-HBsAg-a as the labelled antibody. For clinical application, the new developed diagnostic kit detected HBsAgs of adr, adw, ayr, and ayw at a rate of lower than 0.25, 0.25, 0.5, and 0.5 ng/mL, respectively. Results indicated that this kit was more rapid and sensitive than that other current ELISA-based kits coated with a single MAb (e.g., anti-HBsAg-a).","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 1 1","pages":"47-52"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701300060445","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60498649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.1089/02724570152057661
Kil-Lyong Kim
{"title":"ATOM-2, specific for S-peptide-tagged recombinant proteins.","authors":"Kil-Lyong Kim","doi":"10.1089/02724570152057661","DOIUrl":"https://doi.org/10.1089/02724570152057661","url":null,"abstract":"","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 2 1","pages":"140"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570152057661","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60499689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.1089/027245701300060580
{"title":"4B6 Anti-Dengue-4 Virus, E Recombinant Protein (TERP).","authors":"","doi":"10.1089/027245701300060580","DOIUrl":"https://doi.org/10.1089/027245701300060580","url":null,"abstract":"","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 1 1","pages":"67"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/027245701300060580","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60499162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.1089/02724570152057670
{"title":"MAb to rat galanin.","authors":"","doi":"10.1089/02724570152057670","DOIUrl":"https://doi.org/10.1089/02724570152057670","url":null,"abstract":"","PeriodicalId":55044,"journal":{"name":"Hybridoma","volume":"20 2 1","pages":"141"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/02724570152057670","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60499297","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：