201012929 九、發明說明： 【發明所屬之技術領域】 本發明涉及-種單株抗體，特収關於—種抗流產 披衣菌之單株抗體。 【先前技術】 習知披衣菌（C/z/owjWa/ej)係一種革蘭氏陰性菌， 披衣菌可感染之宿主非常廣泛，該宿主包含人牛、羊、 ❹ 豬、老鼠、魚類及禽類等動物。坡衣菌結構中之外膜蛋白 質（outer membrane protein ; 0MP)具有抗原特性，該外 膜蛋白質係由外膜蛋白3 ( OMP3 )、外膜蛋白2 ( OMP2 ) 、多型性外膜蛋白質（POMP )及主要外膜蛋白質（ MOMP)所組成；其中該主要外膜蛋白質（M〇Mp)佔外 膜蛋白質組成中之60%，使該主要外膜蛋白質（M〇MP )成為披衣菌之重要且具有抗原性的蛋白質，又該主要外 膜蛋白包含VD1、VD2、VD3及VD4四種蛋白，其中該 © VD2分子量大約為22.3 KDa，該VD4分子量大約為 22.7 KDa，且該VD2蛋白為流產披衣菌所獨特之蛋白。 披衣菌係依據其16S及23S核糖體核糖核酸（rRNA )之演化關係進行分類，可將披衣菌分成四科、二屬及九 種’其中該四科係分別為衣原體科（)、副 衣原體科（Parac/i/fl/wjjWiiceize )、怒卡體科（ •SVmhmVzceiie )及華診體科（所)，該二屬係分 别為农琢後屬（Chlamydia)反今衣後M ( Chlamydophila) ’該九種係分別為砂眼披衣菌（inac/iowaiz·5 )、 201012929 猪型披衣菌（C/z/awyi/zV? ίΜΖ*·?)、鼠型披衣菌（ muridarum)、肺炎故衣蛰（Chlamydophila pneumoniae)、 熟轉故衣菌（Chlamydophila psittaci )、雜類坡衣菌（ Chlamydophila felis)、·♦、類坡衣菌（Chlamydophiia caviae )、歌類坡衣菌（CMamycbphila pecorum)及流產彼衣菌 (CMamydoj?hi!a aborts);且各坡衣菌種又可細分為多種 血清型，所以在眾多型別中，將各種坡衣菌作一個區別就 顯的非常重要。 上述披衣菌種中之流產披衣菌係一人畜共通傳染之 病菌’若懷孕之母牛、母羊等反g類動物感染流產披衣菌 ’則容易造成該懷孕之母牛、母羊等反g類動物流產的現 象，而導致畜牧業經濟之嚴重損失；且若人類感染流產披 衣菌’亦會導致人類流產的現象；因而對人類生命造成很 大的威脅。 習知對抗披衣菌之單株抗體，如中華民國專利公告 第181991號「對抗披衣菌之單株抗體及製造該抗體之方 法」’其揭示一種可對抗披衣菌之單株抗體，該習知單株 抗體係由融合瘤TC-19C或融合瘤1〇7_1〇或融合瘤5〇1_ 25所分减造，該習知單株抗體之抗原決定部位係披衣 菌之脂多糖（LPS)上10K、43K或14K的位置。然而， 被衣菌之種類繁多，各種類之披衣菌的基因排列及特性亦 不相同，使各種類之披衣菌的抗原性有所差異，導致該習 知對抗披衣8之單株抗體無法全面性的抵抗每—種類:彼 衣菌’進而無法專-且有效的抵抗上述之流產披衣菌。 201012929 迄今尚未出現可專一又有效對抗流產坡衣菌之單株 抗體。為能有效防治流產披衣菌所造成之經濟損失及人類 生命的咸脅，研發可專一且有效對抗流產彼衣菌之單株抗 體係刻不容緩的課題。 【發明内容】 為了解決流產披衣菌所造成之社會成本損失，本發 明人進行努力研究而研發出一種可專一且有效對抗流產披 衣菌之單株抗體。 本發明之主要目的係提供一種抗流產披衣菌之單株 抗體，使得本發明可專一且有效對抗流產披衣菌。 本發明之另一目的係提供一種可產製抗流產披衣菌 單株抗體之融合瘤細胞株，使得本發明可產製能專一且有 效對抗流產披衣菌之單株抗體。 本發明提供一種抗流產披衣菌之單株抗體及產製該 單株抗體之融合瘤細胞株；該融合瘤細胞株所產製的單株 抗體可與流產披衣菌之主要外膜蛋白質之VD2蛋白專一 性結合，進而藉由動物體内之體液免疫反應消除流產披衣 菌’使得本發明具有可專一防治流產披衣菌之功效。 【實施方式】 為了讓本發明之上述和其他目的、特徵和優點能更 明確被了解，下文將特舉本發明較佳實施例，並配合所附 圖式，作詳細說明如下。 本發明係提供一種可產製專一抗流產披衣菌單株抗 體之融合瘤細胞株’其係利用重組流產披衣菌主要外膜蛋 201012929 白質之VD2蛋白為抗原，對BALB/c mice母系實驗鼠進 行腹腔免疫注射，使該實驗鼠產生高力價抗體反應，進一 步取出實驗鼠之脾臟細胞及骨趙瘤細胞（Ns—〗）進行融 合，並以酵素連結免疫分析法筛選出對流產彼衣菌之外膜 蛋白具有特異反應的融合瘤細胞，將該融合瘤細胞進行單 株化後得到單株融合瘤細胞株，而該融合瘤細胞株所產製 的單株抗體對流產披衣菌主要外膜蛋白具專一性結合反應 〇201012929 IX. Description of the Invention: [Technical Field] The present invention relates to a monoclonal antibody, and a monoclonal antibody against a species of anti-abortion chlamydia. [Prior Art] Traditional chlamydia (C/z/owjWa/ej) is a Gram-negative bacterium, and the host of infection by Chlamydia is very extensive. The host contains human cattle, sheep, pigs, mice, fish. And animals such as poultry. Outer membrane protein (OMP) has antigenic properties in the structure of Chlamydomonas, and the outer membrane protein is composed of outer membrane protein 3 (OMP3), outer membrane protein 2 (OMP2), and polymorphic outer membrane protein (POMP). And the major outer membrane protein (MOMP); wherein the major outer membrane protein (M〇Mp) accounts for 60% of the outer membrane protein composition, making the major outer membrane protein (M〇MP) important for chlamydia And having an antigenic protein, and the main outer membrane protein comprises four proteins VD1, VD2, VD3 and VD4, wherein the VD2 has a molecular weight of about 22.3 KDa, the VD4 has a molecular weight of about 22.7 KDa, and the VD2 protein is aborted. The unique protein of the fungus. Chlamydia is classified according to the evolutionary relationship between its 16S and 23S ribosomal ribonucleic acid (rRNA), which can divide chlamydia into four families, two genera and nine species, of which the four families are Chlamydia () Chlamydia family (Parac/i/fl/wjjWiiceize), anger card body (•SVmhmVzceiie) and Chinese medical department (the institute), the two genus are the genus of the genus Chlamydia, Chlamydophila 'The nine lines are Chlamydia trachomatis (inac/iowaiz·5), 201012929, Chlamydia pneumoniae (C/z/awyi/zV? ΜΖ*??), Chlamydia muridarum, Pneumonia Chlamydophila pneumoniae, Chlamydophila psittaci, Chlamydophila felis, ♦, Chlamydophiia caviae, Chamycbphila pecorum and Amylobacter? (CMamydoj?hi!a aborts); and each species can be subdivided into a variety of serotypes, so in many types, it is very important to make a distinction between the various strains. Among the above-mentioned chlamydia species, the abortive chlamydia is a common infection of the diseased animal. If the pregnant cow, the ewes and other anti-g-like animals are infected with abortion, the cows and ewes are likely to be caused. The phenomenon of abortion of anti-g-type animals leads to serious loss of animal husbandry economy; and if human infection with abortion chlamydia also causes abortion, it poses a great threat to human life. A monoclonal antibody against Chlamydia, such as the Chinese Patent Publication No. 181991 "Single antibody against Chlamydia and a method of producing the same", which discloses a monoclonal antibody against Chlamydia, which The conventional monoclonal antibody system is reduced by the fusion tumor TC-19C or the fusion tumor 1〇7_1〇 or the fusion tumor 5〇1_25, and the epitope of the conventional monoclonal antibody is the lipopolysaccharide of the chlamydia (LPS). ) The position of 10K, 43K or 14K. However, there are many kinds of chlamydia, and the gene arrangement and characteristics of various kinds of chlamydia are also different, which makes the antigenicity of various kinds of chlamydia different, resulting in the conventional antibody against the coat 8 It is impossible to comprehensively resist each type: Pediatrics' and thus cannot specifically and effectively resist the above mentioned abortion chlamydia. 201012929 So far, there has not been a single antibody that is specific and effective against Chlamydia. In order to effectively prevent the economic loss caused by abortion chlamydia and the salty threat of human life, it is an urgent task to develop a single plant resistance system that can specifically and effectively fight against A. faecalis. SUMMARY OF THE INVENTION In order to solve the social cost loss caused by abortion chlamydia, the inventors have diligently researched and developed a monoclonal antibody which is specific and effective against Chlamydia abortus. The main object of the present invention is to provide a monoclonal antibody against Chlamydomonas aeruginosa, so that the present invention can specifically and effectively combat Chlamydia abortus. Another object of the present invention is to provide a fusion tumor cell strain which can produce a monoclonal antibody against Chlamydomonas aeruginosa, so that the present invention can produce a monoclonal antibody which is specific and effective against Chlamydia abortus. The present invention provides a monoclonal antibody against Chlamydomonas aeruginosa and a fusion tumor cell strain producing the monoclonal antibody; the monoclonal antibody produced by the fusion tumor cell line and the main outer membrane protein of Chlamydia abortus The unique combination of VD2 protein and the elimination of abortion chlamydia by humoral immune response in animals makes the invention have the effect of specifically preventing and treating abortion chlamydia. The above and other objects, features and advantages of the present invention will become more <RTIgt; The present invention provides a fusion tumor cell strain capable of producing a single anti-abortion chlamydia monoclonal antibody, which utilizes a VD2 protein of a recombinant abortive chlamydia main outer membrane egg 201012929 white matter as an antigen, and a BALB/c mice maternal experiment. The mice were intraperitoneally injected to induce high-valence antibody reaction, and the spleen cells of the rats and the bone tumor cells (Ns-〗) were further removed for fusion, and the aborted clothes were screened by enzyme-linked immunoassay. A fusion tumor cell having a specific reaction of the membrane protein of the bacterium, and the single-plant fusion tumor cell strain is obtained by monocrocing the fusion tumor cell, and the monoclonal antibody produced by the fusion tumor cell strain is mainly for abortion of the cloaca Outer membrane protein has a specific binding reaction
本發明之流產披衣菌主要外膜蛋白質之VD2蛋白的製備 本聲明之製備流產披衣菌主要外膜蛋白質之VD2蛋 白的第一步驟：係由已感染流產披衣菌之母羊隻的子宮頸 及陰道内採集分泌物檢體，將該分泌物檢體進行漠縮、離 心及聚合酶連敍應（PCR) #實驗流程，叫幅流產披 衣菌主要外膜蛋白基因。本發明之製備流產披衣菌主要外 膜蛋白質之VD2蛋白的第二步驟：係將職產披衣菌主 要外膜蛋白之㈣蛋白基因建構於大轉g表現系統之 載體中’取該載體轉殖人宿主細胞巾，_大量表現流產 披衣菌重組主要㈣蛋自質，㈣成本㈣之流產披衣菌 白質之VD2蛋白的製備。並同步製備流產披 衣菌主要外膜蛋白質之VD4蛋白，以將該彻蛋白做為 對照組並與該㈣蛋白精蛋自質絲讀（sds page I，該蛋白質電^結果如第1圖所示，m為蛋白質標準 :二 *白’Une2為該對照組v_ ,顯不該上清液所呈現之蛋白質分子量為 22.3 KDa 201012929 ，對照組VD4所呈現之分子量為22.7 KDa，由此數據可 確認本發明所製備之VD2蛋白及對照組VD4蛋白正確無 * 誤。 本發明之以流產披衣菌主要外膜蛋白質之VD2蛋白進行 動物免疫試驗 ❹Preparation of the VD2 protein of the major outer membrane protein of Chlamydia abortus of the present invention. The first step of preparing the VD2 protein of the main outer membrane protein of the aborted chlamydia is: the child of the ewes infected with the aborted chlamydia The secretions were collected from the cervix and the vagina, and the secretion samples were subjected to a contraction, centrifugation, and polymerase chain reaction (PCR). The experimental procedure was called the main outer membrane protein gene of the australis. The second step of preparing the VD2 protein of the main outer membrane protein of Chlamydomonas aeruginosa according to the present invention is: constructing the (four) protein gene of the main outer membrane protein of the chlamydia chrysogenum in the vector of the large-g expression system Colonization of host cell towels, _ a large number of abortion recombination of chlamydia mainly (four) egg self-quality, (four) cost (four) of the preparation of VD2 protein of abortion white chlamydia. And simultaneously preparing the VD4 protein of the main outer membrane protein of Chlamydia abortus, using the Schein protein as a control group and reading the (4) protein spermatozoa from the filament (sds page I, the protein is as shown in Fig. 1 It is shown that m is the protein standard: two * white 'Une2 is the control group v_, the protein molecular weight of the supernatant is 22.3 KDa 201012929, and the molecular weight of the control VD4 is 22.7 KDa, the data can be confirmed The VD2 protein prepared by the invention and the VD4 protein of the control group are correct and correct. The VD2 protein of the main outer membrane protein of Chlamydia abortus is used for animal immunity test.
本發明之實驗動物係選自國家實驗研究院之BALB/c mice母系實驗鼠。另外調配一含有流產披衣菌主要外膜 蛋白質之VD2蛋白之注射液，該注射液係包含該VD2蛋 白及弗氏元全佐劑（complete Freund’s adjuvant)，其中該 VD2蛋白及弗氏完全佐劑的體積比為1 : 1，且該蛋 白之》辰度為100//g/ml。本發明之實驗動物免疫試驗的第 一步驟：係將該注射液以腹腔注射之方式施打至實驗鼠體 内以π成第一次免疫，且於第一次免疫後每隔兩週，同樣 以腹腔注射方式進行實驗鼠之補強免疫一次。本發明之實 驗動物免疫試驗的第二步驟：係於進行四次補強免疫後， 以眼窩採血之方式採集實驗鼠血液，並由該血液中取得實 驗鼠進行免賴鎌之血清，該血料有對抗流產披衣菌 抗體，進-步該實驗鼠進行後續之單株抗體製作。 本發明〈可產製抗流產坡衣8之單株抗翻敲合瘤細跑株 本發明之融合瘤細胞株的製備彳法係利用 P〇lyeth却glycol (PEG)進行實驗鼠的骨趙瘤細胞（ Ns-υ及脾臟細胞賴合，再_㈣與單株化 ，以製得可產製抗流產披衣菌之單株抗趙的融合瘤細胞株 201012929 本發明之製備該融合瘤細胞的第一步驟：係取出經 過上述免疫試驗後之實驗鼠的骨髓瘤細胞及脾臟細胞。將 該骨髓瘤細胞以適當的培養基（含5%胎牛血清及L-glutamine的RPMI-1640)進行培養，並培養至該骨趙瘤 細胞數達5x106 /ml。將該免疫試驗後之實驗鼠的脾臟細 胞壓碎，並以RPMI-1640溶液洗滌2次後，將該脾臟細 胞與該骨髓瘤細胞以3 : 1的體積比混合均勻進行融合。 本發明之製備該融合瘤細胞的第二步驟：係將該融合瘤細 胞經 PEG-1500 ( polyethylene glycol-1500 )細胞融合劑處 理後，並以含20%胎牛血清的HAT培養液進行培養，且 在此培養液内唯有骨髓瘤與脾臟細胞融合的細胞才能存活 ，以篩選出融合成功之細胞，並將該融合成功之細胞以 ELISA方法檢測，以由該融合成功之細胞中再篩選出對流 產坡衣菌之外膜蛋白之VD2蛋白具有特異反應的融合瘤 細胞。本發明之製備該融合瘤細胞的第三步驟：係進一步 進行該融合瘤細胞的單株化以獲得單株融合瘤細胞株，而 該融合瘤細胞株所產製的單株抗體對流產披衣菌主要外膜 蛋白之VD2蛋白具專一性結合反應，且也可辨識羊隻胎 盤受流產坡衣菌之感染。 本發明之抗流產披衣菌之單株抗體的製備舆該單株抗艎辨 識流產披衣菌主要外膜蛋白之專一性檢測 將上述所筛選出之融合瘤細胞株進行大量培養，並 取該融合瘤細胞株大量培養後之上清液，該上清液帶有該 201012929 融合瘤細胞株所產製的單株抗體，進一步取該上清液做為 一級抗體，並以 anti-mouse HRP-IgG、IgA 及 IgM 做為二 級抗體以進行流產披衣菌主要膜蛋白之VD2蛋白的西方 轉潰分析及免疫球蛋白亞型鑑定，並同時以VD4蛋白做 為對照組，以碟認該單株抗體可專一性的辨識流產披衣菌 主要外膜蛋白之VD2蛋白。該西方轉潰分析之結果如第 2圖所示，本發明以八株融合成功之融合瘤細胞株進行試 驗’該八株融合瘤細胞株之編號分別為SD-1、SD-2、SD_ 3、SEM、SD_5、SD-6、SD-7 及 SD-8，並以 Anti-His 抗 體辨識VD2蛋白及VD4蛋白以做為控制組，圖中顯示控 制組中之VD2蛋白及VD4蛋白皆產生辨識反應，表示該 VD2蛋白及該VD4蛋白皆可被使用於進行西方轉潰分析 ;由圖中可知，該八株融合瘤細胞株之試驗皆顯示本發明 之融合瘤細胞株所產製的單株抗體，可專一的辨識流產披 衣菌主要膜蛋白之VD2蛋白，而該VD4蛋白並不會被本 發明之融合瘤細胞株所產製的單株抗體所辨識，可證實本 發明之融合瘤細胞株所產製的單株抗體可專一的辨識流產 披衣卤主要膜蛋白之VD2蛋白，進而可藉由動物體内之 體液免疫反應消除流產披衣菌，使得本發明具有可專一防 治流產披衣菌之功效。該免疫球蛋白亞型鑑定之結果如第 3圖所示，免疫球蛋白亞型鑑定主要以酵素連結免疫分析 法測定，以確定本發明之融合瘤細胞株所產製的單株抗體 是屬於 IgGl、IgG2a、IgG2b、IgG3、IgA 及 IgM 免疫球 蛋白中的那一群，又是/c或λ的那一型，該結果顯示本發 201012929 明之融合瘤細胞株所產製的單株抗體是屬於IgG1群及尤 型。 ' 賴該單株抗體可專—性的賴流產披衣菌主要外 膜蛋白之VD2蛋白後，本發明進—步⑽單株抗體辨識 受流產披衣®絲之羊隻胎盤巾㈣產披衣菌，以證實本 發明之抗流產披衣菌之抗體可直接由動物之組織或細麟 識出流產披衣菌之存在；本發明係利用免疫組織化學染色 法(immunochemical staining ; IHC)檢測受流產披衣菌感染 之羊隻胎盤細胞中流產披衣菌的存在，如第4圖之箭頭所 標示，可確認本發明之抗流產披衣菌之單株抗體的確可直 接由動物之組織或細胞辨識出流產披衣菌。 如上所述，相較於習知對抗披衣菌之單株抗體無法 專一抵抗流產披衣菌之缺點，本發明之抗流產披衣菌之單 株抗體可專一的辨識流產披衣菌，以達到可專一抵抗流產 披衣菌之功效。 ◎ 雖然本發明已利用上述較佳實施例揭示，然其並非 用以限定本發明，任何熟習此技藝者，在不脫離本發明之 精神和範圍之内，當可作各種更動與修改，因此本發明之 ‘ 保護範圍當視後附之申請專利範圍所界定者為準。 —12 — 201012929 【圖式簡單說明】 第1圖··本發明之抗流產披衣菌單株抗體之蛋白質電泳 • 圖。 • 第2圖：本發明之抗流產披衣菌單株抗體之西方轉潰分 析結果圖。 第3圖：本發明之抗流產披衣菌單株抗體之酵素連結免 疫分析結果圖。 第4圖：本發明之抗流產披衣菌單株抗體之羊隻胎盤組 ® 織分析結果圖。 ' 【主要元件符號說明】 (無） —13 —The experimental animal of the present invention is selected from the BALB/c mice maternal experimental mouse of the National Experimental Research Institute. Further, an injection comprising a VD2 protein containing a major outer membrane protein of Chlamydia aeruginosa, the injection comprising the VD2 protein and complete Freund's adjuvant, wherein the VD2 protein and Freund's complete adjuvant are formulated The volume ratio is 1:1 and the protein has a degree of 100//g/ml. The first step of the experimental animal immunoassay of the present invention is that the injection is administered intraperitoneally into the test body by π into the first immunization, and every two weeks after the first immunization, the same The vaccination of the experimental mice was performed once by intraperitoneal injection. The second step of the experimental animal immunoassay of the present invention is that after four times of tonifying immunity, the blood of the experimental mouse is collected by blood sampling in the eye socket, and the blood of the test mouse is obtained from the blood, and the blood is obtained. Against the aborted chlamydia antibody, the experimental mouse was further subjected to subsequent monoclonal antibody production. The present invention is capable of producing a single anti-tumor tumor-killing strain of the anti-abortion slope coat 8 of the present invention. The preparation method of the fusion cell line of the invention uses P〇lyeth but glyco (PEG) to perform bone tumor of the experimental mouse. The cells (Ns-υ and spleen cells are ligated, and then _(4) and monocultured to obtain a single anti-Zhao-resistant fusion tumor cell strain capable of producing anti-abortion chlamydia. 201012929 The preparation of the fusion tumor cells of the present invention First step: the myeloma cells and spleen cells of the experimental mice after the above immunoassay are taken out, and the myeloma cells are cultured in an appropriate medium (containing 5% fetal bovine serum and RPMI-1640 of L-glutamine). The number of the tumor cells was 5×106 /ml, and the spleen cells of the experimental mouse after the immunoassay were crushed and washed twice with the RPMI-1640 solution, and the spleen cells and the myeloma cells were 3 The second step of preparing the fusion tumor cell of the present invention is as follows: the fusion tumor cell is treated with PEG-1500 (polyethylene glycol-1500) cell fusion agent, and is contained in 20%. The HAT medium of fetal calf serum is cultured, In this culture, only the cells in which the myeloma and the spleen cells are fused can survive, and the successfully fused cells are selected, and the successfully fused cells are detected by ELISA to be screened out from the successfully fused cells. A fusion tumor cell having a specific reaction to a VD2 protein of a membrane protein of a bacterium of the genus Candida. The third step of preparing the fusion tumor cell of the present invention is to further carry out the singularization of the fusion tumor cell to obtain a single fusion tumor. The cell strain, and the monoclonal antibody produced by the fusion tumor cell strain has a specific binding reaction to the VD2 protein of the main outer membrane protein of the aborted chlamydia, and can also recognize the infection of the sheep placenta by the abortion strain. Preparation of a single antibody against Chlamydia abortus against the invention 舆 Identification of the specific outer membrane protein of Chlamydia aeruginosa by the single plant, and the above-mentioned selected fusion tumor cell strain is cultured in a large amount, and the fusion is taken The supernatant of the tumor cell strain is cultured in a large amount, and the supernatant contains the monoclonal antibody produced by the 201012929 fusion tumor cell line, and the supernatant is further used as the primary antibody. Anti-mouse HRP-IgG, IgA and IgM were used as secondary antibodies to perform Western translocation analysis and immunoglobulin subtype identification of VD2 protein of the main membrane protein of abortive chlamydia, and VD4 protein was used as a control. The group recognizes that the monoclonal antibody specifically recognizes the VD2 protein of the main outer membrane protein of Chlamydia aborigis. The results of the western collapse analysis are shown in Fig. 2, and the present invention has eight fusion fusion tumors. Cell lines were tested. The eight fusion tumor cell lines were numbered SD-1, SD-2, SD_3, SEM, SD_5, SD-6, SD-7 and SD-8, and were identified by Anti-His antibody. VD2 protein and VD4 protein were used as control group. The figure showed that the VD2 protein and VD4 protein in the control group all produced an identification reaction, indicating that the VD2 protein and the VD4 protein can be used for western collapse analysis; It can be seen that the test of the eight fusion tumor cell lines shows that the monoclonal antibody produced by the fusion tumor cell line of the present invention can specifically recognize the VD2 protein of the main membrane protein of the aborted chlamydia, and the VD4 protein does not. Produced by the fusion tumor cell strain of the present invention By identifying the antibody of the strain, it can be confirmed that the monoclonal antibody produced by the fusion tumor cell line of the present invention can specifically recognize the VD2 protein of the main membrane protein of the aborted distillal halogen, and can eliminate the abortion by the humoral immune reaction in the animal. Clostridium, the invention has the effect of specifically preventing and controlling abortion chlamydia. The results of the identification of the immunoglobulin subtype are shown in Fig. 3. The immunoglobulin subtype identification is mainly determined by enzyme-linked immunoassay to determine that the monoclonal antibody produced by the fusion tumor cell line of the present invention belongs to IgGl. The group of IgG2a, IgG2b, IgG3, IgA, and IgM immunoglobulins, which is again the type of /c or λ, and the results show that the monoclonal antibody produced by the fusion tumor cell line of the present invention of 201012929 belongs to IgG1. Group and special type. After the monoclonal antibody can specifically produce the VD2 protein of the main outer membrane protein of chlamydia, the present invention proceeds to step (10) identification of a single antibody to be aborted, and the only placenta of the sheep (four) The bacteria can be used to confirm the presence of the anti-abortion chlamydia antibody of the present invention, and the presence of abortion chlamydia can be directly recognized by the tissue or the stalk of the animal; the present invention detects the abortion by immunochemical staining (IHC). The presence of abortion chlamydia in the placental cells of the chlamydial-infected sheep, as indicated by the arrow in Fig. 4, confirms that the monoclonal antibodies against Chlamydomonas aeruginosa of the present invention can be directly recognized by the tissues or cells of the animal. Abortion of chlamydia. As described above, compared with the conventional single antibody against Chlamydia, the monoclonal antibody against Chlamydia abortus can specifically recognize the abortion chlamydia to achieve It can specifically resist the effect of abortion chlamydia. Although the present invention has been disclosed in the above-described preferred embodiments, it is not intended to limit the invention, and various modifications and changes can be made without departing from the spirit and scope of the invention. The scope of the invention is defined by the scope of the patent application. —12 — 201012929 [Simplified Schematic] Fig. 1 · Protein electrophoresis of a monoclonal antibody against chlamydia in the present invention. • Fig. 2 is a graph showing the results of western collapse analysis of the monoclonal antibodies against Chlamydia pneumoniae of the present invention. Fig. 3 is a graph showing the results of immunoassay analysis of the antibody against a single antibody against Chlamydia abortus of the present invention. Fig. 4 is a graph showing the results of analysis of the sheep placenta group of the anti-abortion chlamydia monoclonal antibody of the present invention. ' [Main component symbol description] (none) —13 —