无机种子颗粒聚集介导的溶菌酶结晶增强,Crystal Growth & Design

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无机种子颗粒聚集介导的溶菌酶结晶增强
Crystal Growth & Design ( IF 3.2 ) Pub Date : 2017-01-30 00:00:00 , DOI: 10.1021/acs.cgd.6b01026
Ulrike Weichsel ₁ , Doris Segets _{1,

2} , Thaseem Thajudeen _{1,

2} , Eva-Maria Maier ₁ , Wolfgang Peukert _{1,

2}

Affiliation

我们表明聚集在蛋白质晶体的种子生长中起主要作用。分批接种的方法通过添加确定量的充分表征的种子颗粒，为设置蛋白质结晶的起始条件提供了机会。对四方鸡蛋蛋白溶菌酶（LSZ）的实验观察证实了较大的结构单元定向聚集以形成蛋白质晶体的概念。结果表明，就更大，更明确的LSZ晶体而言，就加速反应动力学而言，种子颗粒/生物共轭物的聚集对于产品质量而言是有利的。考虑到生长单元的聚集以形成蛋白质晶体，我们提出了LSZ的种子批次结晶的种群平衡（PB）模型。为了模拟晶体生长，通过动态光散射（DLS）测量了结聚的LSZ分子的不断演变的粒度分布（PSD）。此外，通过DLS研究了结晶条件下LSZ溶液中种子颗粒的聚集。符合我们的预期，发现种子的数量很重要，因为它在PB模型的聚集期内会严重影响碰撞频率。最终，所应用的模型正确地给出了过饱和损耗曲线的趋势，以及所测得的CSD粒径的数量级，范围从几纳米到微米大小的颗粒/晶体。因此，通过结合PB模型和实验确定的结晶参数，获得了对晶体形成机理的见解。据我们所知，

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Enhanced Crystallization of Lysozyme Mediated by the Aggregation of Inorganic Seed Particles

We show that aggregation plays a major role in seeded growth of protein crystals. The seeded batch approach provides the opportunity to set the starting conditions for protein crystallization by adding a defined amount of well-characterized seed particles. The experimental observations for tetragonal hen egg-white lysozyme (LSZ) confirm the concept of the oriented aggregation of larger building blocks to form a protein crystal. It was shown that the aggregation of the seed particles/bioconjugates is advantageous for the product quality in terms of larger and more defined LSZ crystals and in terms of accelerated reaction kinetics. We present a population balance (PB) model for the seeded batch crystallization of LSZ considering the aggregation of growth units to form protein crystals. For the modeling of crystal growth, evolving particle size distributions (PSDs) of agglomerating LSZ molecules were measured by dynamic light scattering (DLS). Moreover, the aggregation of seed particles in LSZ solutions under crystallization conditions was investigated by DLS. In line with our expectations, the number of seeds was found to be important as it strongly affects the collision frequency in the aggregation term of our PB model. Finally, the applied model gives trends of the supersaturation depletion curves and orders of magnitude of the measured CSDs in particle size correctly, ranging from only a few nanometers up to micrometer-sized particles/crystals. Thus, by the combination of PB modeling and experimentally determined crystallization parameters, insights into the crystal formation mechanism were obtained. To the best of our knowledge, this is the first attempt to model growth within a crystal population by an aggregation mechanism induced by seeding with foreign particles.

更新日期：2017-01-30

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