Liquid secondary ion mass spectrometric analysis of natural and recombinant proteins and monoclonal antibody light chains with molecular weights between 16 000 and 25 000
Liquid secondary ion mass spectrometric analysis of natural and recombinant proteins and monoclonal antibody light chains with molecular weights between 16 000 and 25 000
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对分子量在 16 000 至 25 000 之间的天然和重组蛋白以及单克隆抗体轻链进行液体二次离子质谱分析
DOI：
10.1021/ac00213a039
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发表时间：
1990
影响因子：
7.4
通讯作者：
I. Hollander
中科院分区：
文献类型：
--
作者：
M. Siegel;R. Tsao;V. Doelling;I. Hollander
最近，Lacey和Keough（I）总结了分子量大于15000的小蛋白质的质谱数据。用于这些测量的电离方法是等离子体解吸（PD），电喷雾，激光解吸，和液体SIMS（二次离子质谱）。液体二次离子质谱的结果是基于Barber和绿色（2）对小模型蛋白的原始工作。Barber和绿色通过使用铯离子枪（在35 kV下操作）演示了液体SIMS技术，并获得了单电荷和多电荷分子离子。在最近的工作（3）中，使用猪胰蛋白酶（MW 23460）作为模型化合物，高质量液相-SIMS谱的两个特征被证明，即，（1）信号随样品量线性增加，直至在0.6 nM（14 n%）下饱和，信噪比（S/N）为5，和（2）S/N降低，当样品中掺入100倍摩尔过量的氯化钠时，N增加约2.5倍（60 nM氯化钠对应于1.4 μ g氯化钠）。对于低分子量化合物（< 3000）也观察到饱和和掺杂效应。然而，在16000和25000道尔顿之间的质量下，注意到小的S/N动态范围和少量掺杂物对信号的剧烈影响。在本文中，我们将详细描述获得有用的液体二次离子质谱数据的单一组分的中等纯度和简单的混合物，分子量在16000和25000之间的实验条件。研究的小蛋白质是天然和重组生长激素，重组生长因子，并减少单克隆抗体的轻链。这些化合物的液体SIMS数据将与更传统的生化方法获得的结果进行比较。这些蛋白质具有农业或药用价值。生长激素和生长因子用于农业和药物化学中以促进生长，抗体用于免疫靶向研究。
Recently, Lacey and Keough (I) summarized mass spectral data obtained for small proteins with molecular weights greater than 15000. The ionization methods used for these measurements were plasma desorption (PD), electrospray, laser desorption, and liquid-SIMS (secondary ion mass spectrometry). The liquid-SIMS results were based upon the original work of Barber and Green (2) on small model proteins. Barber and Green demonstrated the liquid-SIMS technique by using a cesium ion gun, operating at 35 kV, and obtained singly as well as multiply charged molecular ions. In more recent work (3), using porcine trypsin (MW 23460) as the model compound, two features of high mass liquid-SIMS spectra were demonstrated, namely,(1) the linear increase in signal with sample size until saturation at 0.6 nM (14 n%) with a signal-to-noise (S/N) ratio of 5 and (2) the decrease in S/N by about a factor of 2.5 times when the sample was adulterated with 100 times molar excess of NaCl (60 nM of NaCl corre-sponds to 1.4 tig of NaCl). Saturation and adulteration effects have also been observed for low molecular weight compounds (< 3000). However, at masses between 16000 and 25000 daltons one notes the small S/N dynamic range and the drastic effect on the signal by small amounts of adulterant. In this paper, we will describe in detail the experimental conditions necessary for obtaining useful liquid-SIMS data on single components of moderate purity and simple mixtures with molecular weights between 16000 and 25000. The small proteins studied were natural and recombinant somatotropins, recombinant growth factors, and reduced light chains of monoclonal antibodies. The liquid-SIMS data for these compounds will be compared with results obtained by more traditional biochemical methods. These proteins are of ag-ricultural or pharmaceutical interest. The somatotropins and growth factors are used in agricultural and medicinal chemistry for growth promotion and the antibodies are used in immunological targeting studies.
DOI：
10.1073/pnas.84.5.1127
发表时间：
1987-03-01
影响因子：
11.1
作者：
RECSEI, PA;GRUSS, AD;NOVICK, RP
通讯作者：
NOVICK, RP