公开(公告)号：KR100665054B1

公开(公告)日：2007-01-04

申请号：KR1020040083994

申请日：2004-10-20

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  고영준 ,  나진주 ,  조남인

IPC: C12N15/79 ,  C12N15/63

CPC classification number: C12N7/00 ,  A61K2039/5258 ,  C12N2770/32323

Abstract: 본 발명은 유전자 재조합 기법을 이용한 돼지 수포병 바이러스 유사입자(Swine Vesicular Disease Virus-Like Particle) 및 그 제조 방법에 관한 것으로, 보다 상세하게는 돼지 수포병 바이러스 전체 유전자 중에서 구조단백질 전구체(P1) 유전자 및 3CD 프로테아제(protease) 유전자를 베큘로 바이러스 발현벡터에 동시 삽입하여 곤충세포에서 발현함으로써, 발현된 3CD 프로테아제 효소가 구조단백질 전구체를 개별적인 단백질로 절단하여 개별단백질들이 자체적인 조립과정을 통해서 돼지 수포병 바이러스의 캡시드(capsid)를 형성하는 것을 특징으로 하는 돼지 수포병 바이러스 유사입자의 제조 방법에 관한 것이다.
유전자 재조합, 돼지 수포병 바이러스 유사입자, 진단용 항원, 백신개발, 구조단백질 전구체, 3CD 프로테아제

公开(公告)号：KR1020060040879A

公开(公告)日：2006-05-11

申请号：KR1020040089816

申请日：2004-11-05

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  나진주 ,  고영준 ,  조남인

IPC: G01N33/68 ,  G01N33/53

CPC classification number: G01N33/56983 ,  G01N2469/20 ,  C07K16/1027 ,  C07K2317/14 ,  C07K2319/00 ,  C07K2319/21 ,  C12N5/163 ,  C12N15/866 ,  G01N33/577 ,  G01N2333/115

Abstract: The present invention relates to a diagnostic method of Peste des Petits Ruminants (PPR), a viral animal disease. More specifically, the invention relates to a new diagnostic method capable of detecting PPRV antibodies safely, simply and rapidly with recombinant Nucleocapsid (N) protein of PPR virus (PPRV) and specific monoclonal antibody against N protein from PPRV.

公开(公告)号：KR100430934B1

公开(公告)日：2004-05-14

申请号：KR1020010082976

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  현방훈 ,  고영준 ,  이광녕 ,  구복경 ,  엄재구 ,  나진주 ,  남향미 ,  박지용 ,  계수정 ,  김인중 ,  이세영 ,  최광석 ,  손현주 ,  주이석

IPC: C12N15/33

Abstract: PURPOSE: Diagnostic methods of foot-and-mouth disease using recombinant 3ABC non-structural protein expressed in insect cells and a monoclonal antibody is provided, thereby more rapidly and accurately diagnosing the foot-and-mouth disease than the prior methods. CONSTITUTION: A gene encoding a recombinant 3ABC non-structural protein derived from Korean foot-and-mouth disease virus O/SKR/2000 has the nucleotide sequence of SEQ ID NO: 1. A recombinant baculovirus to be expressed in insect cells is prepared by co-transfection with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. The recombinant 3ABC non-structural protein is expressed by infection of the insect cells with the recombinant baculovirus. A hybridoma cell line 3F-11(KCTC 10138BP) is prepared by introducing the recombinant 3ABC non-structural protein expressed in E. coli into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A monoclonal antibody is produced from the hybridoma cell line 3F-11(KCTC 10138BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant 3ABC non-structural protein in a coating buffer solution and pouring the diluate on a plate; (2) reacting the testing serum with the plate; (3) reacting a conjugate, which binds with an antibody for foot-and-mouth disease virus in the testing serum and has an enzyme, a radioactive material or a fluorescent material, with the testing serum; and (4) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.

Abstract translation: 目的：提供使用在昆虫细胞和单克隆抗体中表达的重组3ABC非结构蛋白的口蹄疫诊断方法，从而比现有方法更快速和准确地诊断口蹄疫。 构成：编码来源于韩国口蹄疫病毒O / SKR / 2000的重组3ABC非结构蛋白的基​​因具有SEQ ID NO：1的核苷酸序列。在昆虫细胞中表达的重组杆状病毒由 用含有SEQ ID NO：1的重组3ABC非结构蛋白基因的重组载体共转染。通过用重组杆状病毒感染昆虫细胞表达重组3ABC非结构蛋白。 通过将在大肠杆菌中表达的重组3ABC非结构蛋白导入动物中，从动物中收集免疫的细胞并将免疫的细胞与癌细胞融合来制备杂交瘤细胞系3F-11（KCTC 10138BP）。 从杂交瘤细胞系3F-11（KCTC 10138BP）产生单克隆抗体。 一种口蹄疫诊断方法，包括以下步骤：（1）将重组3ABC非结构蛋白稀释于包被缓冲液中，并将稀释液倒入平板中; （2）使测试血清与平板反应; （3）使与试验血清中的口蹄疫病毒抗体结合的缀合物与测试血清具有酶，放射性物质或荧光物质; 和（4）测量酶反应的强度，荧光反应或与缀合物的辐射反应。

公开(公告)号：KR1020040022247A

公开(公告)日：2004-03-12

申请号：KR1020010082714

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  권병준 ,  고영준 ,  이세영 ,  김인중 ,  주이석

IPC: A61K39/12

Abstract: PURPOSE: A method of diagnosing vesicular stomatitis by performing an ELISA method using the recombinant envelope glycoprotein antigen of vesicular stomatitis virus(VSV) and monoclonal antibodies thereon is provided. Therefore, it permits quick, exact and specific antibody detection of the VSV and thus exact diagnosis of vesicular stomatitis through the detection. CONSTITUTION: The diagnosing method for vesicular stomatitis is achieved through a VSV detection method consisting of: attaching monoclonal antibodies against an envelope glycoprotein antigen of VSV on a solid supporter; binding the envelope glycoprotein antigen to the monoclonal antibodies attached on the solid supporter in the first step; reacting a sample containing an antibody against the envelope glycoprotein antigen with the envelope glycoprotein antigen of the second step; reacting a secondary antibody selected from the group consisting of a conjugate antibody attached with enzyme and a conjugate antibody attached with a luminescent material with the antibody of the third step.

Abstract translation: 目的：提供使用水泡性口炎病毒（VSV）的重组包膜糖蛋白抗原及其单克隆抗体进行ELISA方法诊断水泡性口炎的方法。 因此，它可以快速，精确和特异性地检测VSV，从而通过检测确定水泡性口炎的诊断。 构成：水泡性口炎的诊断方法是通过VSV检测方法实现的，该方法包括：将固定支持物上VSV的包膜糖蛋白抗原的单克隆抗体连接起来; 在第一步中将包膜糖蛋白抗原与附着在固体支持物上的单克隆抗体结合; 使含有针对包膜糖蛋白抗原的抗体的样品与第二步的包膜糖蛋白抗原反应; 使选自由酶附着的结合抗体和附着有发光材料的缀合物抗体的第二抗体与第三步骤的抗体反应。