山羊皮革膠原蛋白快速檢測試紙的制備及應(yīng)用

摘 要： 為檢測古代皮革文物，制備了一種山羊皮革膠原蛋白快速檢測試紙。通過設(shè)計(jì)制備膠原蛋白多克隆抗體，并采用膠體金作為示蹤物對(duì)抗體進(jìn)行標(biāo)記，利用免疫反應(yīng)完成后產(chǎn)生的顏色變化檢測山羊皮革膠原蛋白；優(yōu)化膠體金標(biāo)記抗體的最適pH和最佳抗體標(biāo)記量，利用不同濃度的山羊皮革膠原蛋白溶液分析了試紙的靈敏度，選用7種抗原分析了試紙的特異性。結(jié)果表明：在膠體金標(biāo)記抗體的pH值8、抗體標(biāo)記量為48 μg/mL時(shí)，所制備的快速檢測試紙對(duì)山羊皮革膠原蛋白的檢出限（Limit of detection， LOD）可達(dá)800 ng/mL，15 min內(nèi)可獲得檢測結(jié)果，并且具有優(yōu)異的特異性，能有效排除一些干擾蛋白的影響，精準(zhǔn)鑒別山羊皮制文物。該研究可為古蛋白文物考古現(xiàn)場快速檢測研究提供新思路。

關(guān)鍵詞：古代皮革文物；膠原蛋白多克隆抗體；膠體金；快速檢測試紙；考古現(xiàn)場

中圖分類號(hào)：TS195.644

文獻(xiàn)標(biāo)志碼：A

文章編號(hào)：1673-3851 （2025） 03-0214-07

引文格式：徐冰，李藝暢，張超，等. 山羊皮革膠原蛋白快速檢測試紙的制備及應(yīng)用［J］. 浙江理工大學(xué)學(xué)報(bào)（自然科學(xué)），2025，53（2）：214-220.

Reference Format： XU Bing，LI Yichang，ZHANG Chao，et al. Preparation and application of an assay strip for the rapid detection of collagen in goat leather［J］. Journal of Zhejiang Sci-Tech University，2025，53（2）：214-220.

Preparation and application of an assay strip for the rapid detection of collagen in goat leather

XU Bing， LI Yichang， ZHANG Chao， WANG Bing

（College of Materials Science amp; Engineering， Zhejiang Sci-Tech University， Hangzhou 310018， China）

Abstract：To detect ancient leather artifacts， a test strip for rapid collagen detection in goat leather was prepared. The collagen polyclonal antibody was prepared by design， and colloidal gold was used as a tracer to label the antibody， and the colour change produced after the completion of the immune reaction was used to detect goat leather collagen. The optimum pH and the optimum amount of colloidal gold-labelled antibody were optimized， the sensitivity of the test strip was analyzed by using different concentrations of goat leather collagen solutions， and the specificity of the test strip was analyzed by choosing seven kinds of antigens. The results showed that when the pH value of colloidal gold-labeled antibody was 8 and the labelled amount of antibody was 48 μg/mL， the limit of detection （LOD） of the prepared rapid detection test strip for goat leather collagen could reach 800 ng/mL， and the detection results could be obtained within 15 min. The test strip had excellent specificity， could effectively eliminate the influence of some interfering proteins and accurately identify goat skin artifacts. This study offers a new approach for the rapid detection of ancient protein artifacts at archaeological sites.

Key words：ancient leather artifacts; collagen polyclonal antibody; colloidal gold; rapid test strip; archaeological site

0 引 言

在絲、麻、棉等纖維材料出現(xiàn)之前，皮革就已廣泛應(yīng)用于人類的服飾^［1］。《禮記·禮運(yùn)》記載了古代人們“未有絲麻，衣其羽皮”的情形。《后漢書·輿服志》中也提到，“上古穴居而野處，衣毛而冒皮”。然而，由于受到埋藏環(huán)境的長期影響，在光、熱、水汽和微生物等作用下，大多數(shù)皮革文物在出土?xí)r就已腐爛不堪，失去了原有的形態(tài)^［2-3］。

傳統(tǒng)的皮革文物鑒定方法主要包括光學(xué)顯微鏡、掃描電子顯微鏡、傅里葉變換紅外光譜和核磁共振波譜法等^［4］。……