生物制剂和生物仿制药的特征

Biologics and biosimilars, also known as biotherapeutic drugs, are produced from living organisms such as bacteria, yeast and mammalian cells: this is in contrast to small molecule drugs, which are synthesised chemically. This means that on top of being very large molecules (peptides, small proteins, antibodies, polysaccharides etc.), they exhibit post-translational modifications and certain degrees of structure variability.

Confirmation of the identity of the therapeutic drug through structural characterisation, from the primary amino acid sequence to the higher order structure, together with impurity control are critical factors to ensure efficacy and patients safety.

布鲁克的质谱解决方案超出了传统的肽映射和完整的质量分析。这些解决方案将最终的性能，速度和多功能性与强大的功能流动性软件相结合，可为生物制剂开发人员提供正交表征方法：从单克隆抗体（MAB）亚基分析使用中间和中间的方法，用于序列的无迹象检测和表征变体和修饰，包括脱氨酸，氧化和N末端剪辑到高阶结构研究，以详细表征二硫键的详细表征，构象表位，在抗体药物结合物（ADC）中扰动以计算准确的药物抗体比率（DAR）的结构变化。

Disulfide bond (DSB) analysis and hydrogen deuterium exchange (HDX) have emerged as technologies that can be employed to gain insights into protein structure. These insights are critical for determining structural similarity for biosimilars, or for monitoring protein stability during drug development. The tertiary structure of therapeutic proteins is key to their activity and stability. 

研发实验室需要基于未还原蛋白质的单一摘要，并且在没有酶特异性或天然DSB的事先了解的情况下，可以在生物药物中自动化DSB分析。由于这些蛋白质的复杂性以及它们将包含多种二硫键键的事实，分析是一个挑战，通常需要通过减少和未还原蛋白质的胰蛋白酶消化以及对这两种分析的手动比较进行多次LC-MS运行。

NMR is especially sensitive to changes to higher order structure at atomic resolution, making it ideally suited for similarity assessment of biologics and biosimilars (high order structure). NMR allows for intact protein analysis, enabling evaluation of the structure of therapeutic drugs without modification, in conditions that are physiologically relevant. Due to the quantitative nature of magnetic resonance and its selectivity, potency determination, impurity profiling and degradation studies (e.g. 多山矿石) are performed directly enabling fast and easy testing without the need of response factor calculations, or the method redevelopment activities required by traditional LC methods, thereby saving time and reducing costs. In addition, benchtop time domain NMR (TD-NMR) is becoming an important technique for the analysis of蛋白质聚集体and 100%fill-check小瓶和注射器。

傅里叶变换红外(ir)光谱n be used to analyse water-soluble and membrane proteins such as nuclear receptors, which are currently a very important targets in drug research and development, being associated with conditions such as Alzheimer, Parkinson diseases, diabetes and obesity. FT-IR is typically applied in protein secondary structure elucidation, detection of conformational changes and monitoring of protein dynamics like aggregation, precipitation and crystallization. Infrared protein analysis is a quick and relatively inexpensive technique used for formulation optimisation, stability studies during drug development and QC of protein drug products.

表面等离子体共振（SPR）是从动力学角度表征相互作用的黄金标准方法。尽管基于解决方案的方法为相互作用提供了基于平衡的观点，但SPR的无标签和实时分析可提供对动力学的见解，从而增加了一个关键特征，以更精确地了解交互作用。在基于平衡的方法（例如ELISA）中具有相似亲和力的生物制剂可能具有不同的非速率，随后具有不同的生物学结果。因此，SPR可以在目标蛋白质上使用最佳半衰期鉴定生物制剂。

SPR进一步允许鉴定靶向蛋白质上不同表位的抗体。同样，有条件的结合基于pH，例如就像在癌症组织中一样，或可以轻松研究不同缓冲液对结合相互作用的影响。这些方法有助于理解在生理上更可翻译的环境中的互动。