本文關(guān)鍵詞：抗軟骨藻酸單克隆抗體制備及ELISA檢測(cè)方法建立　出處：《吉林大學(xué)》2008年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 軟骨藻酸 單克隆抗體 快速免疫 ELISA

【摘要】： 軟骨藻酸由海洋中的硅藻產(chǎn)生后常通過(guò)食物鏈蓄積于海洋貝類(lèi)、蛤類(lèi)等海產(chǎn)品體內(nèi),進(jìn)而引起海洋動(dòng)物乃至人的食物中毒,國(guó)外由于誤食有毒海產(chǎn)品而引發(fā)中毒的事件時(shí)有發(fā)生,嚴(yán)重影響了人們的身體健康和對(duì)海產(chǎn)品的開(kāi)發(fā)利用。本實(shí)驗(yàn)通過(guò)活潑酯法將軟骨藻酸同牛血清白蛋白及人免疫球蛋白進(jìn)行偶聯(lián),成功制備了免疫原及檢測(cè)原,通過(guò)足墊快速法免疫BALB/C小鼠,常規(guī)方法進(jìn)行融合,間接ELISA方法篩選陽(yáng)性克隆,同時(shí)采用有限稀釋法、顯微操作法及皮下荷瘤方法進(jìn)行雜交瘤細(xì)胞的亞克隆和培養(yǎng),建立了9C1雜交瘤細(xì)胞株,并對(duì)其單克隆抗體的效價(jià)、分子量、抗體亞類(lèi)、有效濃度、染色體眾數(shù)、純度、蛋白質(zhì)含量、親和力、特異性等方面進(jìn)行了研究,利用優(yōu)化后的實(shí)驗(yàn)條件進(jìn)行間接競(jìng)爭(zhēng)ELISA檢測(cè)并建立標(biāo)準(zhǔn)曲線,標(biāo)準(zhǔn)曲線的回歸方程為y = -13.094x + 106.19,相關(guān)系數(shù)R2=0.9962,最低檢測(cè)限2.43ng/mL。
[Abstract]:After being produced by diatoms in the ocean, alginic acid often accumulates in marine shellfish, clams and other marine products through the food chain, which leads to food poisoning of marine animals and even people. Poisoning has occurred from time to time in foreign countries due to the accidental consumption of toxic seafood. In this experiment, the alginic acid was coupled with bovine serum albumin and human immunoglobulin by active ester method, and the immunogen was prepared successfully. BALB/C mice were immunized with foot pad fast method, fusion was performed by routine method, positive clones were screened by indirect ELISA method, and limited dilution method was used at the same time. The hybridoma cell line 9C1 was subcloned and cultured by micromanipulation and subcutaneous tumor-bearing method. The monoclonal antibody titer, molecular weight, antibody subclass and effective concentration of the monoclonal antibody were obtained. Chromosome mode, purity, protein content, affinity, specificity and other aspects were studied. The optimized experimental conditions were used for indirect competitive ELISA detection and the establishment of standard curve. The regression equation of the standard curve is y = -13.094x 106.19, the correlation coefficient R _ 2N _ 2 is 0.9962and the minimum detection limit is 2.43ng / mL.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類(lèi)號(hào)】：R155.3;R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前3條

1 高利利;程金平;劉元Z，

本文編號(hào)：1423981