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公开(公告)号:US4792601A
公开(公告)日:1988-12-20
申请号:US656482
申请日:1984-10-01
IPC分类号: C07K16/00 , C07K16/18 , C12N5/16 , G01N33/53 , A61K39/395 , C12N5/00 , C12N15/00 , C12P21/00
摘要: In a method for preparing a monoclonal hybridoma which secretes antibodies against an immunogenic material, comprising sensitizing an appropriate source with an immunizing amount of a preparation of the immunogenic material, obtaining from the source lymphocytes having immune activity against the immunogenic material, and fusing the lymphocytes with an appropriate cell line to thereby form mixed hybridomas, the improvement wherein: the immunogenic material is present in the immunizing preparation in an amount not larger than 10% by weight, and wherein, after formation of the mixed hydridomas but prior to cloning, the method comprises the steps of: raising ascites fluid with the mixed hydridomas, then detecting immune specificity against the immunogenic material in the ascites raised fluid; and thereafter cloning the ascites raised cells which show immune specificity against the material.
摘要翻译: 在制备分泌针对免疫原性物质的抗体的单克隆杂交瘤的方法中,包括用免疫量的免疫原性物质的制剂对适当的来源进行敏化,从源获得具有针对免疫原性物质的免疫活性的淋巴细胞,并融合淋巴细胞 用合适的细胞系形成混合的杂交瘤,其改进在于:免疫原性物质以不大于10重量%的量存在于免疫制剂中,并且其中,在形成混合的hydridomas之后但是在克隆之前, 方法包括以下步骤:用混合的hydridomas提升腹水,然后检测腹水引起的流体中对免疫原性物质的免疫特异性; 然后克隆显示对材料的免疫特异性的腹水升高的细胞。
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2.
公开(公告)号:US4487833A
公开(公告)日:1984-12-11
申请号:US353089
申请日:1982-03-01
摘要: In a method for preparing a monoclonal hybridoma which secretes antibodies against an immunogenic material, comprising sensitizing an appropriate source with an immunizing amount of a preparation of the immunogenic material, obtaining from the source lymphocytes having immune activity against the immunogenic material, and fusing the lymphocytes with an appropriate cell line to thereby form mixed hybridomas, the improvement wherein: the immunogenic material is present in the immunizing preparation in an amount not larger than 5% by weight, and wherein, after formation of the mixed hybridomas but prior to cloning, the method comprises the steps of: raising ascites fluid with the mixed hybridomas, then detecting immune specificity against the immunogenic material in the ascites raised fluid; and thereafter cloning the ascites raised cells which show immune specificity against the material.
摘要翻译: 在制备分泌针对免疫原性物质的抗体的单克隆杂交瘤的方法中,包括用免疫量的免疫原性物质的制剂对适当的来源进行敏化,从源获得具有针对免疫原性物质的免疫活性的淋巴细胞,并融合淋巴细胞 用适当的细胞系形成混合的杂交瘤,其改进在于:免疫原性物质以不超过5%的量存在于免疫制剂中,并且其中,在形成混合杂交瘤之后但是在克隆之前, 方法包括以下步骤:用混合杂交瘤提升腹水,然后检测腹水引起的流体中对免疫原性物质的免疫特异性; 然后克隆显示对材料的免疫特异性的腹水升高的细胞。
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3.
公开(公告)号:US4404188A
公开(公告)日:1983-09-13
申请号:US287943
申请日:1981-07-29
CPC分类号: A61K38/22 , B01D15/362 , B01D15/363 , B01D15/1871 , Y10S530/85
摘要: A process for purifying Mullerian Inhibiting Substance (MIS) from testicular tissue which comprises incubating testicular tissue in a culture media in the presence of a protease inhibitor in order to extract MIS from the tissue, chromatographing the extract on an anionic exchange resin followed by chromatographing the biologically active product obtained therefrom on a cationic exchange resin; the biologically active material from the cationic exchange resin is chromatographed by lectin affinity chromatography on wheat germ lectin and then either chromatographed by lectin affinity chromatography on concanavalin A or by triazinyl dye affinity chromatography on an appropriate triazinyl dye; MIS is obtained in 8,000-fold and 15,000-fold purification over original testicular tissue.
摘要翻译: 一种从睾丸组织中纯化Mullerian抑制物质(MIS)的方法,包括在蛋白酶抑制剂存在下在培养基中孵育睾丸组织以从组织中提取MIS,在阴离子交换树脂上色谱分离提取物,随后色谱分离 由阳离子交换树脂得到的生物活性产物; 阳离子交换树脂的生物活性物质通过凝集素亲和层析对小麦胚芽凝集素进行色谱分离,然后通过凝血素亲和层析对刀豆蛋白A进行色谱分离,或通过三嗪基染料亲和色谱法在合适的三嗪基染料上进行色谱分离; 在原始睾丸组织中获得了8000倍和15000倍的纯化的MIS。
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公开(公告)号:US4510131A
公开(公告)日:1985-04-09
申请号:US512553
申请日:1983-07-11
CPC分类号: A61K38/22 , Y10S530/828 , Y10S530/85
摘要: A process for purifying Mullerian Inhibiting Substance (MIS) from testicular tissue which comprises incubating testicular tissue in a culture media in the presence of a protease inhibitor in order to extract MIS from the tissue, chromatographing the extract on an anionic exchange resin followed by chromatographing the biologically active product obtained therefrom on a cationic exchange resin; the biologically active material from the cationic exchange resin is chromatographed by lectin affinity chromatography on wheat germ lectin and then either chromatographed by lectin affinity chromatography on concanavalin A or by triazinyl dye affinity chromatography on a appropriate triazinyl dye; MIS is obtained in 8,000-fold and 15,000-fold purification over original testicular tissue.
摘要翻译: 一种从睾丸组织中纯化Mullerian抑制物质(MIS)的方法,包括在蛋白酶抑制剂存在下在培养基中孵育睾丸组织以从组织中提取MIS,在阴离子交换树脂上色谱分离提取物,随后色谱分离 由阳离子交换树脂得到的生物活性产物; 阳离子交换树脂的生物活性物质通过凝集素亲和层析对小麦胚芽凝集素进行色谱分离,然后通过凝血素亲和层析对刀豆蛋白A进行色谱分离,或通过三嗪基染料亲和色谱法在合适的三嗪基染料上进行色谱分离; 在原始睾丸组织中获得了8000倍和15000倍的纯化的MIS。
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