Novel Laser-Based Deposition of Active Protein Thin Films
Novel Laser-Based Deposition of Active Protein Thin Films
复制标题
活性蛋白薄膜的新型激光沉积
作者：
B. Ringeisen;J. Callahan;P. K. Wu;A. Piqué;B. Spargo;R. McGill;M. Bucaro;Heungsoo Kim;and D. M. Bubb;D. Chrisey
本文报道了一种新的基于激光的沉积活性蛋白质薄膜的方法，即基质辅助脉冲激光蒸发(Maple)。我们已经均匀地沉积了10 nm到近1μm的胰岛素和辣根过氧化物酶薄膜。我们进行了几个实验来表征沉积薄膜的化学完整性。在枫树沉积的胰岛素膜上进行的基质辅助激光解吸/电离和液相色谱/电喷雾电离质谱学实验表明，这种沉积技术中涉及的激光−材料相互作用并不改变蛋白质的质量。傅里叶变换红外光谱实验表明，枫树沉积的HRP的化学功能和二级结构与天然蛋白质几乎相同。我们还发现，沉积的HRP膜保持了催化3，3‘-二氨基联苯胺(DAB)还原的能力，这表明转移蛋白的活性部位不受活性中心的影响。
This paper reports the deposition of active protein thin films by a novel laser-based approach termed matrix-assisted pulsed laser evaporation (MAPLE). We have deposited uniform 10 nm to nearly 1 μm thin films of insulin and horseradish peroxidase (HRP). We performed several experiments to characterize the chemical integrity of the deposited films. Matrix assisted laser desorption/ionization and liquid chromatography/electrospray ionization mass spectrometry experiments performed on MAPLE-deposited insulin films indicate that the laser−material interaction involved in this deposition technique does not modify the protein's mass. Fourier transform infrared spectroscopy experiments show that the chemical functionality and secondary structure of MAPLE-deposited HRP are nearly identical to those of the native protein. We also find that deposited HRP films retain their ability to catalyze the reduction of 3,3‘-diaminobenzidine (DAB), suggesting that the active site of transferred proteins is unaffected by the ...