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三點(diǎn)鑒定抗血清質(zhì)量

時(shí)間:2011/11/1閱讀:934
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 抗血清的質(zhì)量直接影響分析方法的特異性和靈敏度。用于鑒定抗血清質(zhì)量的參數(shù)主要有幾項(xiàng): 
    1.親和性(affinity)  是指抗體分子上一個(gè)抗原結(jié)合部位與相應(yīng)的抗原決定簇之間的結(jié)合強(qiáng)度,常用親和常數(shù)Ka表示。Ka是正/逆向反應(yīng)速度常數(shù)的比值,單位為稀釋度單位(L/mol或LM-1),表示需將lmol抗體稀釋至多少升,才能使抗原—抗體結(jié)合下降50%;而Ka的倒數(shù)為解離常數(shù)(Kd),其單位為濃度單位(mol/L)。為提高放射免疫分析的靈敏度、精密度和準(zhǔn)確度,需選用Ka值高(109一1012L/mo1)的抗血清。
     2.特異性(specificity)  是指一種抗體識別相應(yīng)抗原決定簇的能力。由于一種抗原常有結(jié)構(gòu)極相似的類似物(同類物、前體、降解物和代謝物),其抗血清可能對該抗原的類似物產(chǎn)生交叉反 應(yīng)。因此,抗體的特異性通常以交叉反應(yīng)率表示:將反應(yīng)的zui大結(jié)合率抑制下降50%時(shí)特異性抗原與類似物的劑量(ED50)之比來計(jì)算交叉反應(yīng)率。如胰島素標(biāo)準(zhǔn)品使125I胰島素與抗血清的zui大結(jié)合下降50%的用量為0.5ng,而類似的物(前體)胰島素原的用量為500ng,則相應(yīng)的交叉反應(yīng)率(%)為:0.5/500=0.1%。
抗體交叉反應(yīng)程度的高低直接影響測定結(jié)果的準(zhǔn)確性,但應(yīng)注意,抗血清特異性與分析方法特異性并不*相同,因后者對某一待測抗原的特異性程度不僅取決于所用抗體,尚有賴于標(biāo)記物的質(zhì)量和方法學(xué)設(shè)計(jì)。
     3.效價(jià)(titer)  是反映抗血清中有效抗體含量的相對參數(shù),即抗血清稀釋至能與抗原發(fā)生有效反應(yīng)的zui大稀釋度。通常是將一株抗血清作系列稀釋后與標(biāo)記抗原反應(yīng),計(jì)算不同稀釋 度時(shí)抗體結(jié)合標(biāo)記抗原的百分率,繪制出抗體稀釋度曲線。結(jié)合具體方法設(shè)計(jì)要求。效價(jià)一般指結(jié)合50%標(biāo)記抗原時(shí),抗血清的稀釋度。
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