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    • 33. 发明公开
    • 돼지오제스키바이러스에 대한 단일클론 항체와 이를 생산하는 하이브리도마 세포주 및 상기 단일클론 항체를 이용한 돼지오제스키 항체 검출법
    • 针对AUJESZKY病毒的MONOCLONAL抗体及其生产的HYBRIDOMA细胞系以及AUJESZKY抗体检测方法
    • KR1020010039001A
    • 2001-05-15
    • KR1019990047205
    • 1999-10-28
    • 주식회사 메디안디노스틱
    • 차명진박흥숙장병식
    • C07K16/08
    • Y02E10/20F03B17/02F03B13/06Y02E10/223
    • PURPOSE: Provided are a monoclonmal antibody against Aujeszky virus and a hybridoma cell line producing the same. And a detection method of Aujeszky antibody using the monoclonal antibody is also provided. CONSTITUTION: Aujeszky virus monoclonal antibody and its activity are detected by the steps of: (i) inoculating a swine kidney cell with a mouse antigen, culturing it and followed by inactivating culture supernatant and inoculating a swine kidney cell with it to prepare antogenic protein for immunization; (ii) mixing the antigenic protein with immuno adjuvant and followed by injecting it into a mouse and collecting inguinal lymphnode from the mouse; (iii) culturing myeloma cell in DMEM medium containing BFS to prepare myeloma cell line; (iv) preparing a hybridoma cell line by fusing inguinal lymphnode with myeloma cell line; (v) absorbing inactivated Aujeszky virus and a contrast antigen on ELISA microplate and followed by reacting it with hybridoma cell culture supernatant and antimouse HRP conjugate and adding chromogen ABTS to select a cell line that produces moclonal antibody specific to Aujeszky virus; (vi) injecting the monoclonal antibody into mouse abdomen to produce abdominal dropsy then followed by purifying the monoclonal antibody therefrom, concentrating and conjugating it with peroxidase; (vii) dividing inactivated Aujeszky virus on microplate and followed by reacting it with bovine serum albumin, transplanting onto ELISA plate and reacting it with the conjugate; (viii) adding chromogen ABTS and measuring optical density to detect monoclonal antibody and its activity.
    • 目的:提供针对Aujeszky病毒的单克隆抗体和产生该抗体的杂交瘤细胞系。 还提供了使用单克隆抗体的Aujeszky抗体的检测方法。 构成:通过以下步骤检测Aujeszky病毒单克隆抗体及其活性:(i)用小鼠抗原接种猪肾细胞,培养它,然后灭活培养上清液并用其接种猪肾细胞以制备用于 免疫接种; (ii)将抗原蛋白与免疫佐剂混合,然后将其注入小鼠并从小鼠收集腹股沟淋巴结; (iii)在含有BFS的DMEM培养基中培养骨髓瘤细胞以制备骨髓瘤细胞系; (iv)通过将腹股沟淋巴结与骨髓瘤细胞系融合来制备杂交瘤细胞系; (v)在ELISA微量培养板上吸收灭活的Aujeszky病毒和对比抗原,随后将其与杂交瘤细胞培养上清液和抗小鼠HRP缀合物反应,并加入染色体ABTS以选择产生针对Aujeszky病毒的特异性抗体的细胞系; (vi)将单克隆抗体注射入小鼠腹部产生腹水,然后纯化单克隆抗体,浓缩并与过氧化物酶缀合; (vii)将灭活的Aujeszky病毒分离在微量培养板上,随后将其与牛血清白蛋白反应,移植到ELISA板上并与缀合物反应; (viii)加入色原ABTS并测量光密度以检测单克隆抗体及其活性。
    • 34. 发明授权
    • B형간염바이러스의다른두항원을동시에인식하는이중특이항체및그의제조방법
    • 乙型肝炎病毒BISPECIFIC抗体及其制备方法
    • KR100267058B1
    • 2000-09-15
    • KR1019970076922
    • 1997-12-29
    • 한국과학기술연구원
    • 홍효정류춘제박성섭
    • C07K16/08
    • PURPOSE: A bispecific antibody being capable of recognizing antigens of hepatitis B virus and a producing method thereof are provided, which can recognize hepatitis B virus antigens which are different each other, and is thus effectively used in the prevention and treatment of hepatitis. CONSTITUTION: The bispecific antibody TBSIg recognizing antigens of hepatitis B virus contains amino acid sequence represented by sequence ID No. 12 and comprises a S antigen of hepatitis B virus recognizing region, link peptides, a pre-S2 of hepatitis B virus recognizing region, and human antibody Fc region. An expression vector pCMV-dhfr-TBSIg contains the bispecific TBSIg gene represented by sequence ID No. 11. The transformant E. coli(KCTC 8861P) is produced by transforming with the expression vector pCMV-dhfr-TBSIg. The CHO cell line BSG-18H produces the bispecific antibody TBSIg. The method for producing the bispecific antibody TBSIg comprises the steps of: transforming CHO cell line with the expression vector pCMV-dhfr-TBSIg; selecting CHO cell line capable of mass producing the bispecific antibody by incubating the transformed CHO cell line in a medium containing MTX; transferring the CHO cell line into a serum free medium and incubating it; and subjecting the fermented culture to column chromatography to obtain the bispecific antibody. The sequences ID. No. 11 and 12 above are described as in the description.
    • 目的:提供能够识别乙型肝炎病毒抗原的双特异性抗体及其制备方法,其可以识别彼此不同的乙型肝炎病毒抗原,因此有效地用于预防和治疗肝炎。 构成:识别乙型肝炎病毒抗原的双特异性抗体TBSIg含有由序列号12表示的氨基酸序列,包含乙型肝炎病毒识别区域的S抗原,链接肽,乙型肝炎病毒识别区域的前S2位点,以及 人抗体Fc区。 表达载体pCMV-dhfr-TBSIg含有由序列号11表示的双特异性TBSIg基因。通过用表达载体pCMV-dhfr-TBSIg转化产生转化体大肠杆菌(KCTC 8861P)。 CHO细胞系BSG-18H产生双特异性抗体TBSIg。 制备双特异性抗体TBSIg的方法包括以下步骤:用表达载体pCMV-dhfr-TBSIg转化CHO细胞系; 选择能够通过在含有MTX的培养基中培养转化的CHO细胞系来大量产生双特异性抗体的CHO细胞系; 将CHO细胞系转移到无血清培养基中并孵育; 并将发酵的培养物进行柱色谱以获得双特异性抗体。 序列ID。 上述11和12描述如下。