利用单克隆抗体-胶体金缀合物对杨木纤维素上里氏木霉纤维二糖水解酶 I 和内切葡聚糖酶 I 的可视化。
Visualization of Trichoderma reesei Cellobiohydrolase I and Endoglucanase I on Aspen Cellulose by Using Monoclonal Antibody-Colloidal Gold Conjugates.
机构信息
Biotechnology Research Branch, Fuels & Chemicals Research and Engineering Division, Solar Energy Research Institute, 1617 Cole Blvd., Golden, Colorado 80401, and Departments of Microbiology and Anatomy and Neurobiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado 80523.
出版信息
Appl Environ Microbiol. 1991 Nov;57(11):3163-70. doi: 10.1128/aem.57.11.3163-3170.1991.
Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH I) and endoglucanase I (EG I) were conjugated to 10- and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I was visualized by utilizing the MAb-colloidal gold probes. The visualization procedure involved immobilization of cellulose microfibrils on copper electron microscopy grids, incubation of the cellulose-coated grids with cellulase(s), binding of MAb-colloidal gold conjugates to cellulase(s), and visualization via transmission electron microscopy. CBH I was seen bound to apparent crystalline cellulose as well as apparent amorphous cellulose. EG I was seen bound extensively to apparent amorphous cellulose with minimal binding to crystalline cellulose.
分别将针对纤维二糖水解酶 I（CBH I）和内切葡聚糖酶 I（EG I）的单克隆抗体（MAbs）与 10- 和 15-nm 胶体金颗粒偶联。通过利用 MAbs-胶体金探针来可视化 CBH I 和 EG I 的结合。可视化程序包括将纤维素微纤维固定在铜电子显微镜网格上，用纤维素酶孵育纤维素包被的网格，MAb-胶体金缀合物与纤维素酶结合，然后通过透射电子显微镜进行可视化。可以看到 CBH I 与明显的结晶纤维素以及明显的无定形纤维素结合。EG I 广泛地与明显的无定形纤维素结合，与结晶纤维素的结合很少。
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