Whole serum BSA antibody screening using a label-free biophotonic nanoparticle array.
Whole serum BSA antibody screening using a label-free biophotonic nanoparticle array.
复制标题
使用无标记生物光子纳米颗粒阵列筛选全血清 BSA 抗体。
DOI：
10.1016/j.ab.2008.10.042
复制
发表时间：
2009
影响因子：
2.9
通讯作者：
A. M. Shaw
中科院分区：
文献类型：
--
作者：
R. Olkhov;Jeremy D Fowke;A. M. Shaw
用生物光子阵列技术从兔抗血清中筛选出牛血清白蛋白抗体（aBSA）。该阵列最初用种子金纳米颗粒印刷成96点配置，并在每个点的表面上原位合成130 nm的金纳米颗粒。然后用蛋白质牛血清白蛋白（BSA）、纤维蛋白原和免疫球蛋白G（IgG）以及氨基酸甘氨酸将金纳米颗粒表面官能化。使用来自纳米颗粒的时间依赖性光散射的动力学分析来确定全血清中aBSA的浓度。由该阵列得到的aBSA-BSA动力学参数为ka=（1.3±0.3）× 10 - 5 M-1 s-1，kd=（4±2）×10 - 4s-1，KD= 3 nM，与连续金表面的结果相当。阵列读出器对体折射率（RI）的最终灵敏度为1×10− 4折射率单位（RIU），对应于aBSA的1μgml− 1。纳米颗粒似乎比连续的金表面对来自全血清的aBSA结合事件更敏感，并且这是根据等离子体场中RI对比度的差异来解释的。
Bovine serum albumin antibodies (aBSA) have been screened from whole leporine anti serum on a biophotonic array. The array was initially printed with seed gold nanoparticles into a 96-spot configuration, and 130-nm gold nanoparticles were synthesised in situ on the surface of each spot. The gold nanoparticle surface was then functionalized with the proteins bovine serum albumin (BSA), fibrinogen, and immunoglobulin G (IgG) and with the amino acid glycine. The concentration of aBSA in the whole serum was determined using a kinetic analysis of the time-dependent light scattering from the nanoparticles. The aBSA–BSA kinetic parameters derived from the array are ka=(1.3±0.3)×105M−1s−1, kd=(4±2)×10−4s−1, and KD=3nM, which compare favorably with those from continuous gold surfaces. The ultimate sensitivity of the array reader to the bulk refractive index (RI) is 1×10−4refractive index units (RIU), corresponding to 1μgml−1for aBSA. The nanoparticles appear to be more sensitive than the continuous gold surface to the aBSA binding event from whole serum, and this is interpreted in terms of the difference in RI contrast in the plasmon fields.