BACKGROUND & AIMS: :Plant cells are ideal bioreactors for the production and oral delivery of vaccines and biopharmaceuticals, eliminating the need for expensive fermentation, purification, cold storage, transportation and sterile delivery. Plant-made vaccines have been developed for two decades but none has advanced beyond Phase I. However, two plant-made biopharmaceuticals are now advancing through Phase II and Phase III human clinical trials. In this review, we evaluate the advantages and disadvantages of different plant expression systems (stable nuclear and chloroplast or transient viral) and their current limitations or challenges. We provide suggestions for advancing this valuable concept for clinical applications and conclude that greater research emphasis is needed on large-scale production, purification, functional characterization, oral delivery and preclinical evaluation.

背景与目标： 植物细胞是用于生产和口服递送疫苗和生物制药的理想生物反应器，无需昂贵的发酵，纯化，冷藏，运输和无菌递送。植物疫苗已经开发了二十年，但没有一个疫苗已经超过了I期。但是，目前有两种植物生物药物正在通过II期和III期人体临床试验。在这篇综述中，我们评估了不同植物表达系统（稳定的核和叶绿体或瞬时病毒）的优缺点及其当前的局限性或挑战。我们提供了一些建议，以推动这一有价值的概念在临床上的应用，并得出结论，需要在大规模生产，纯化，功能表征，口服给药和临床前评估方面给予更多的研究重点。

BACKGROUND & AIMS: :It was clear from the number of attendees and the broad spectrum of organizations represented at this conference, that effective formulation development for biopharmaceuticals is a critical and timely issue. From DNA-based products to antibodies, vaccines to therapeutics, administered by every route known to medicine, formulation plays a critical role in the ultimate success of biopharmaceutical products. It was obvious from the various case studies presented that effective formulation development is not often achieved as an organization enters into drug development. In fact, systematic pre-formulation work was stressed by most speakers as an effective use of company resources. In order to develop successful formulations, it is essential that appropriate analytical information is collected, that excipients are systematically evaluated, and that the developer understands the basis of stability of the drug product. In all cases, this means understanding product instability, including the physicochemical and thermodynamic basis for instability. Drugs such as therapeutic antibodies, which may require large doses to achieve the desired pharmacological effect, require ever more advanced formulation development to achieve concentrations in the 100-mg/ml range. Potent drugs, such as gene therapies and DNA vaccines, require formulation development in order to achieve effective delivery and expression. On aggregate, it is apparent that there is much to learn in formulation development, from each other as well as in the unexplored territory ahead. This conference provided an excellent forum for the exchange of ideas and information regarding effective formulation development for biopharmaceuticals.

背景与目标： ：从与会的人数和参加此次会议的众多组织中可以明显看出，有效开发生物制药制剂是一个关键而及时的问题。从基于DNA的产品到抗体，从疫苗到治疗剂，通过医学上已知的每种途径进行给药，制剂在生物制药产品的最终成功中都起着至关重要的作用。从提出的各种案例研究中可以明显看出，随着组织参与药物开发，往往无法实现有效的制剂开发。实际上，大多数发言者都强调系统化的预制定工作是对公司资源的有效利用。为了开发成功的制剂，必须收集适当的分析信息，对赋形剂进行系统评估，并且开发人员必须了解药品稳定性的基础，这一点至关重要。在所有情况下，这都意味着要了解产品的不稳定性，包括不稳定性的物理化学和热力学基础。可能需要大剂量才能达到所需药理作用的药物（例如治疗性抗体），需要开发更先进的制剂才能达到100 mg / ml的浓度。强有力的药物，例如基因疗法和DNA疫苗，需要开发配方才能实现有效的传递和表达。总体而言，很明显，在配方开发方面，彼此之间以及在未来的未开发领域中，还有很多值得学习的东西。这次会议为交流有关生物药物有效制剂开发的思想和信息提供了一个极好的论坛。

BACKGROUND & AIMS: :This Special issue, Biopharmaceuticals - discovery, development and manufacturing is edited by Alois Jungbauer and Klaus Graumann and includes articles on various aspects of the development of biopharmaceuticals. Topics covered include the importance of antigen presentation in creating and selecting binding molecules, G-protein-coupled receptors as drug targets, and continuous downstream processing.

背景与目标： ：本期特刊《生物制药-发现，开发和制造》由Alois Jungbauer和Klaus Graumann编辑，其中包括有关生物制药开发各个方面的文章。涵盖的主题包括抗原呈递在创建和选择结合分子，G蛋白偶联受体作为药物靶标以及持续下游加工中的重要性。

BACKGROUND & AIMS: :Biopharmaceuticals (BPs) represent a rapidly growing class of approved and investigational drug therapies that is contributing significantly to advancing treatment in multiple disease areas, including inflammatory and autoimmune diseases, genetic deficiencies and cancer. Unfortunately, unwanted immunogenic responses to BPs, in particular those affecting clinical safety or efficacy, remain among the most common negative effects associated with this important class of drugs. To manage and reduce risk of unwanted immunogenicity, diverse communities of clinicians, pharmaceutical industry and academic scientists are involved in: interpretation and management of clinical and biological outcomes of BP immunogenicity, improvement of methods for describing, predicting and mitigating immunogenicity risk and elucidation of underlying causes. Collaboration and alignment of efforts across these communities is made difficult due to lack of agreement on concepts, practices and standardized terms and definitions related to immunogenicity. The Innovative Medicines Initiative (IMI; www.imi-europe.org), ABIRISK consortium [Anti-Biopharmaceutical (BP) Immunization Prediction and Clinical Relevance to Reduce the Risk; www.abirisk.eu] was formed by leading clinicians, academic scientists and EFPIA (European Federation of Pharmaceutical Industries and Associations) members to elucidate underlying causes, improve methods for immunogenicity prediction and mitigation and establish common definitions around terms and concepts related to immunogenicity. These efforts are expected to facilitate broader collaborations and lead to new guidelines for managing immunogenicity. To support alignment, an overview of concepts behind the set of key terms and definitions adopted to date by ABIRISK is provided herein along with a link to access and download the ABIRISK terms and definitions and provide comments (http://www.abirisk.eu/index_t_and_d.asp).

背景与目标： 生物药物（BPs）代表了快速增长的一类经过批准和研究的药物疗法，为促进多种疾病领域的治疗做出了重要贡献，包括炎性和自身免疫性疾病，遗传缺陷和癌症。不幸的是，对BP的有害的免疫原性应答，尤其是那些影响临床安全性或功效的免疫原性应答，仍然是与这一重要药物类别相关的最常见的负面作用。为了管理和减少不良免疫原性的风险，临床医生，制药行业和学术界科学家参与了以下工作：解释和管理BP免疫原性的临床和生物学结果，改进描述，预测和减轻免疫原性风险的方法以及阐明潜在的免疫原性。原因。由于缺乏对与免疫原性有关的概念，实践以及标准化术语和定义的共识，使得跨这些社区的工作难以进行协作和统一。创新药物倡议（IMI； www.imi-europe.org），ABIRISK联盟[抗生物制药（BP）免疫预测和降低临床风险的临床意义； www.abirisk.eu]由领先的临床医生，学术科学家和EFPIA（欧洲制药工业和协会联合会）成员组成，旨在阐明根本原因，改进免疫原性预测和缓解方法，并围绕与免疫原性相关的术语和概念建立通用定义。预计这些努力将促进更广泛的合作，并为管理免疫原性带来新的指导方针。为了支持一致性，此处提供了ABIRISK迄今为止采用的一组关键术语和定义背后的概念概述，以及访问和下载ABIRISK术语和定义并提供注释的链接（http://www.abirisk.eu /index_t_and_d.asp）。

BACKGROUND & AIMS: BACKGROUND:The COVID-19 pandemic had infected more than 3.5M people around the world and more than 250K people died in 187 countries by May 2020. The causal agent of this disease is a coronavirus whose onset of symptoms to death range from 6 to 41 days with a median of 14 days. This period is dependent on several factors such as the presence of comorbidities, age and the efficiency of the innate or adaptive immune responses. The effectors mechanisms of both types of immune responses depend on the pathogen involved. In the case of a viral infection, the innate immune response may approach the harmful virus through pattern recognition receptors inducing an antiviral state. On the other hand, the adaptive immune response activates antibody production to neutralize or eliminate the virus. Phenolics are plant secondary metabolites with many biological activities for plants and humans against infection. Chemical modification of proteins may enhance their biological properties; thus, a protein of medical interest, for instance, a viral protein can be used as scaffold to build a biopharmaceutical conjugated or complexated with phenolics exhibiting structural complexity or biological activities to achieve effective phenolic-protein-based therapeutics like vaccine adjuvant complexes, immunogen conjugates, and antiviral conjugates. CONCLUSION:Pharmaceutical biotechnology applies the principles of biotechnology to develop biopharmaceuticals for protein-based therapeutics; such as adjuvants, recombinant proteins, monoclonal antibodies, and antivirals. As neither a vaccine nor a treatment for COVID-19 is currently available, this manuscript focuses on insights from pharmaceutical biotechnology into phenolic biopharmaceuticals against COVID-19.

背景与目标： 背景：到2020年5月，COVID-19大流行已感染了全球超过350万人，并在187个国家中超过25万人死亡。该病的病因是冠状病毒，其症状发作至死亡的范围为6至41。天，中位数为14天。这个时期取决于多种因素，例如合并症的存在，年龄以及先天性或适应性免疫反应的效率。两种类型的免疫反应的效应子机制均取决于所涉及的病原体。在病毒感染的情况下，先天的免疫反应可能通过诱导抗病毒状态的模式识别受体接近有害病毒。另一方面，适应性免疫应答激活抗体产生以中和或消除病毒。酚类物质是植物的次生代谢产物，对植物和人类具有抵抗感染的许多生物学活性。蛋白质的化学修饰可以增强其生物学特性；因此，医学上感兴趣的蛋白质，例如病毒蛋白质可以用作支架，以构建与酚类化合物结合或复合的生物药物，所述酚类化合物表现出结构复杂性或生物学活性，以实现有效的基于酚类蛋白质的疗法，如疫苗佐剂复合物，免疫原结合物和抗病毒偶联物。
结论：药物生物技术应用生物技术原理开发基于蛋白质的治疗药物。例如佐剂，重组蛋白，单克隆抗体和抗病毒药。由于目前尚无针对COVID-19的疫苗或治疗方法，因此该手稿着重介绍了从药物生物技术到针对COVID-19的酚类生物药物的见解。

BACKGROUND & AIMS: :Although toxicology studies should always be conducted in pharmacologically relevant species, the specificity of many biopharmaceuticals can present challenges in identification of a relevant species. In certain cases, that is, when the clinical product is active only in humans or chimpanzees, or if the clinical candidate is active in other species but immunogenicity limits the ability to conduct a thorough safety assessment, alternative approaches to evaluating the safety of a biopharmaceutical must be considered. Alternative approaches, including animal models of disease, genetically modified mice, or use of surrogate molecules, may improve the predictive value of preclinical safety assessments of species-specific biopharmaceuticals, although many caveats associated with these models must be considered. Because of the many caveats that are discussed in this article, alternative approaches should only be used to evaluate safety when the clinical candidate cannot be readily tested in at least one relevant species to identify potential hazards.

背景与目标： ：尽管毒理学研究应始终在药理学上相关的物种中进行，但许多生物药物的特异性可能会给鉴定相关物种带来挑战。在某些情况下，即当临床产品仅在人类或黑猩猩中具有活性，或者如果临床候选者在其他物种中具有活性，但是免疫原性限制了进行彻底安全性评估的能力，则可以使用替代方法来评估生物药品的安全性必须考虑。替代方法，包括疾病的动物模型，转基因小鼠或替代分子的使用，可以提高对物种特异性生物药物进行临床前安全性评估的预测价值，尽管必须考虑与这些模型相关的许多警告。由于本文讨论了许多警告，因此仅当无法在至少一个相关物种中对临床候选者进行便捷测试以识别潜在危害时，才应使用替代方法来评估安全性。

BACKGROUND & AIMS: :Despite the wide occurrence of crystallization in the pharmaceutical industry, deep understanding and fine control of the process remain a tricky issue. Nevertheless, the successful manufacturing of finished pharmaceutical products, as well as the structural determination of biopharmaceuticals, depend on the size, form, shape and purity of the crystals. The ability of substrates with precise chemistry and topological features to induce nucleation has been thoroughly assessed during the recent years. This paper reviews the major advances and discoveries in controlling small molecule drug and protein crystallization by means of engineered surfaces. By designing superficial properties and morphology, it has been possible to tune the polymorph outcome, shorten the nucleation induction time, impose specific crystal shapes, control the crystal size and carry out crystallization at very low supersaturation levels. Such achievements underline the potential of surface-induced crystallization to provide an ideal platform for the study of the nucleation process and gain control over its stochastic nature.

背景与目标： ：尽管在制药行业中发生了广泛的结晶，但是对过程的深入了解和精细控制仍然是一个棘手的问题。然而，成品药品的成功生产以及生物药品的结构确定取决于晶体的大小，形式，形状和纯度。近年来，已经对具有精确化学和拓扑特征的底物诱导成核的能力进行了全面评估。本文综述了通过工程表面控制小分子药物和蛋白质结晶的主要进展和发现。通过设计表面性质和形态，可以调节多晶型物的结果，缩短成核诱导时间，施加特定的晶体形状，控制晶体尺寸并以非常低的过饱和水平进行结晶。这些成就强调了表面诱导结晶的潜力，为研究成核过程和获得对其随机性的控制提供了理想的平台。

BACKGROUND & AIMS: :PEGylation (the covalent binding of one or more polyethylene glycol molecules to another molecule) is a technology frequently used to improve the half-life and other pharmaceutical or pharmacological properties of proteins, peptides, and aptamers. To date, 11 PEGylated biopharmaceuticals have been approved and there is indication that many more are in nonclinical or clinical development. Adverse effects seen with those in toxicology studies are mostly related to the active part of the drug molecule and not to polyethylene glycol (PEG). In 5 of the 11 approved and 10 of the 17 PEGylated biopharmaceuticals in a 2013 industry survey presented here, cellular vacuolation is histologically observed in toxicology studies in certain organs and tissues. No other effects attributed to PEG alone have been reported. Importantly, vacuolation, which occurs mainly in phagocytes, has not been linked with changes in organ function in these toxicology studies. This article was authored through collaborative efforts of industry toxicologists/nonclinical scientists to address the nonclinical safety of large PEG molecules (>10 kilo Dalton) in PEGylated biopharmaceuticals. The impact of the PEG molecule on overall nonclinical safety assessments of PEGylated biopharmaceuticals is discussed, and toxicological information from a 2013 industry survey on PEGylated biopharmaceuticals under development is summarized. Results will contribute to the database of toxicological information publicly available for PEG and PEGylated biopharmaceuticals.

背景与目标： PEG化（一种或多种聚乙二醇分子与另一种分子的共价结合）是一种经常用于改善蛋白质，肽和适体的半衰期以及其他药物或药理特性的技术。迄今为止，已经批准了11种PEG化生物药物，并且有迹象表明，还有更多的药物正在非临床或临床研究中。在毒理学研究中看到的不良反应主要与药物分子的活性部分有关，与聚乙二醇（PEG）无关。在此处呈现的2013年行业调查中，在11种已批准的11种PEG化生物药物中有5种在17种药物中，有10种在组织学上在某些器官和组织的毒理学研究中观察到了细胞空泡化。尚无单独归因于PEG的其他影响的报道。重要的是，在这些毒理学研究中，主要发生在吞噬细胞中的空泡化与器官功能的变化没有联系。本文是通过行业毒理学家/非临床科学家的共同努力撰写的，旨在解决PEG化生物药物中大PEG分子（> 10千道尔顿）的非临床安全性的问题。讨论了PEG分子对PEG化生物药品的整体非临床安全性评估的影响，并总结了2013年有关正在开发的PEG化生物药品行业调查的毒理学信息。结果将有助于公开可用于PEG和PEG化生物制药的毒理学信息数据库。

BACKGROUND & AIMS: :Subcutaneous (SC) injection is becoming a more common route for the administration of biopharmaceuticals. Currently, there is no reliable in vitro method that can be used to anticipate the in vivo performance of a biopharmaceutical formulation intended for SC injection. Nor is there an animal model that can predict in vivo outcomes such as bioavailability in humans. We address this unmet need by the development of a novel in vitro system, termed Scissor (Subcutaneous Injection Site Simulator). The system models environmental changes that a biopharmaceutical could experience as it transitions from conditions of a drug product formulation to the homeostatic state of the hypodermis following SC injection. Scissor uses a dialysis-based injection chamber, which can incorporate various concentrations and combinations of acellular extracellular matrix (ECM) components that may affect the release of a biopharmaceutical from the SC injection site. This chamber is immersed in a container of a bicarbonate-based physiological buffer that mimics the SC injection site and the infinite sink of the body. Such an arrangement allows for real-time monitoring of the biopharmaceutical within the injection chamber, and can be used to characterize physicochemical changes of the drug and its interactions with ECM components. Movement of a biopharmaceutical from the injection chamber to the infinite sink compartment simulates the drug migration from the injection site and uptake by the blood and/or lymph capillaries. Here, we present an initial evaluation of the Scissor system using the ECM element hyaluronic acid and test formulations of insulin and four different monoclonal antibodies. Our findings suggest that Scissor can provide a tractable method to examine the potential fate of a biopharmaceutical formulation after its SC injection in humans and that this approach may provide a reliable and representative alternative to animal testing for the initial screening of SC formulations.

背景与目标： ：皮下（SC）注射正在成为生物药物给药的一种更为普遍的途径。当前，没有可靠的体外方法可用于预期用于SC注射的生物药物制剂的体内性能。也没有一种动物模型可以预测体内的结果，例如人类的生物利用度。我们通过开发称为Scissor（皮下注射部位模拟器）的新型体外系统来满足这一未满足的需求。该系统对生物药物在从SC注射后从药物制剂的状态转变为皮下组织的稳态状态时可能经历的环境变化进行建模。 Scissor使用基于透析的注射腔室，该腔室可以合并各种浓度和脱细胞细胞外基质（ECM）成分的组合，这些成分可能会影响生物药物从SC注射部位的释放。该腔室浸入一个基于碳酸氢盐的生理缓冲液的容器中，该缓冲液模仿了SC注射部位和身体的无限深处。这样的布置允许对注射室内的生物药物进行实时监控，并且可以用于表征药物的物理化学变化及其与ECM成分的相互作用。生物药物从注射室向无限的水槽隔室的运动模拟了药物从注射部位的迁移以及血液和/或淋巴毛细血管的吸收。在这里，我们介绍了使用ECM元素透明质酸和胰岛素以及四种不同单克隆抗体的测试制剂对Scissor系统的初步评估。我们的发现表明，剪刀式剪可提供一种易于处理的方法来检查生物药物制剂在人体中的SC注射后的命运，并且这种方法可为动物试验中SC制剂的初步筛选提供可靠且有代表性的替代方法。

BACKGROUND & AIMS: :The freezing step plays a central role in reaching the most stringent requirements of quality, homogeneity and standardization of freeze-dried products. In this paper, a systematic procedure has been proposed to obtain a quantitative estimation of the pore-size variability of lyophilized products resulting from uncontrollable variations of the nucleation temperature. This procedure consisted in collecting the nucleation temperature from a statistically significant number of samples and correlating each nucleation temperature to the corresponding product morphology, using a mathematical model, to obtain a statistical description of the lyophilized product structure. This approach can also be used to obtain an estimation of the variability of the mass transfer resistance to vapor flow and, finally, of the drying time. Two different freezing configurations, i.e., conventional and suspended-vial freezing, have been used as case studies since they can produce significantly different freezing rates.

背景与目标： ：冷冻步骤在达到冷冻干燥产品的质量，均质性和标准化最严格的要求中起着核心作用。在本文中，已经提出了一种系统的程序来获得对由于成核温度的不可控制的变化而导致的冻干产品的孔径变化的定量估计。该程序包括从统计学上大量的样品中收集成核温度，并使用数学模型将每个成核温度与相应的产品形态相关联，以获得冻干产品结构的统计描述。该方法也可以用于获得传质阻力对蒸气流的变化以及最终的干燥时间的变化的估计。案例研究使用了两种不同的冷冻配置，即常规冷冻和小瓶冷冻，因为它们可以产生明显不同的冷冻速率。

BACKGROUND & AIMS: :Antiphospholipid syndrome (APS) is an autoimmune disease that is characterized by arterial and/or venous thrombosis and/or recurrent pregnancy losses. Obstetric APS (OAPS) is considered as a distinct entity from vascular APS (VAPS). In the absence of any additional disease, APS is designated as primary (PAPS), while the term secondary APS (SAPS) is used when other diseases are associated. Catastrophic APS (CAPS) is characterized by the rapid development of multiple thrombosis in various vital organs. The presence of antiphospholipid antibodies (aPL Abs) is considered as a laboratory criterion for APS diagnosis. aPL Abs cause an increase in systemic and decidual TNF-alpha levels in experimental model of APS (eAPS), while paradoxically, administration of TNF-alpha blockers has been associated with de novo synthesis of aPL Abs in patients with various autoimmune diseases. While eAPS provides evidence for the fact that application of TNF-alpha blockers has beneficial effects, lack of randomized prospective studies is the main obstacle for consideration of TNF-alpha blockers administration as a therapeutic option not for all, but at least for selected cases of APS patients despite compelling evidence for detrimental roles of TNF-alpha for both VASP and OAPS. This article represents a review of previously published reports on detrimental roles of TNF-alpha in APS, reports on the application of anti-TNF-alpha agents in eAPS and articles that reported de novo synthesis of aPL Abs induced by biopharmaceuticals against TNF-alpha.

背景与目标： ：抗磷脂综合症（APS）是一种自身免疫性疾病，其特征是动脉和/或静脉血栓形成和/或反复流产。产科APS（OAPS）被认为是与血管APS（VAPS）不同的实体。在没有任何其他疾病的情况下，将APS称为原发性（PAPS），而在与其他疾病相关的情况下使用术语继发性APS（SAPS）。灾难性APS（CAPS）的特征在于各种重要器官中多发性血栓形成的快速发展。抗磷脂抗体（aPL Abs）的存在被认为是诊断APS的实验室标准。在APS（eAPS）实验模型中，aPL Abs导致全身和蜕膜TNF-α水平增加，而自相矛盾的是，在患有各种自身免疫性疾病的患者中，TNF-α阻滞剂的给药与aPL Abs的从头合成有关。尽管eAPS为使用TNF-α阻滞剂产生有益作用这一事实提供了证据，但缺乏随机的前瞻性研究是考虑将TNF-α阻滞剂作为治疗选择的主要障碍，但并非对所有患者（至少对于某些病例）尽管有令人信服的证据表明APS患者对VASP和OAPS都具有TNF-α的有害作用。本文代表对先前发表的有关TNF-α在APS中的有害作用的报道的综述，有关抗TNF-α药剂在eAPS中的应用的报道，以及报道了由生物药物诱导的针对TNF-α的aPL Ab的从头合成的文章。

BACKGROUND & AIMS: :The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. Glycans can influence ligand binding, serum half-life as well as biological activity or product immunogenicity. Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems.

背景与目标： ：治疗性糖蛋白（包括凝血因子，抗体，细胞因子和激素）市场是生物制药行业中盈利，快速增长且充满挑战的行业之一。尽管哺乳动物细胞培养仍然昂贵并且技术上复杂，但是产生期望的翻译后修饰，特别是糖基化的能力是主要问题。聚糖可影响配体结合，血清半衰期以及生物学活性或产物免疫原性。为了建立重组糖蛋白的新型生产平台，研究了人TE671细胞系。由于对细胞膜蛋白的初步分析显示，出于生物技术目的，TE671细胞有希望的糖基化作用，因此我们集中于两种具有治疗意义的模型糖蛋白的重组表达。人丝氨酸蛋白酶抑制剂α-1-抗胰蛋白酶（A1AT）和造血细胞因子粒细胞巨噬细胞集落刺激因子（GM-CSF）的细胞转染程序和无血清表达的优化成功。通过基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱对两种纯化蛋白进行N-聚糖分析，为了解TE671糖基化潜力提供了基础知识。除了蛋白质特异的模式，还根据各自的TE671细胞培养的培养基条件，观察到了很强的区别-特别是N-聚糖岩藻糖基化和唾液酸化。已经显示出细胞系合成复杂和高度唾液酸化的N-聚糖结构的能力。我们的结果证明TE671细胞系可作为其他现有人类表达系统的替代品。

BACKGROUND & AIMS: :Monoclonal antibodies (mAbs) are an important class of therapeutic agents approved for the therapy of many types of malignancies. However, in certain cases applications of conventional mAbs have several limitations in anticancer immunotherapy. These limitations include insufficient efficacy and adverse effects. The antigen-binding fragments of antibodies have a considerable potential to overcome the disadvantages of conventional mAbs, such as poor penetration into solid tumors and Fc-mediated bystander activation of the immune system. Fragments of antibodies retain antigen specificity and part of functional properties of conventional mAbs and at the same time have much better penetration into the tumors and a greatly reduced level of adverse effects. Recent advantages in antibody engineering allowed to produce different types of antibody fragments with improved structure and properties for efficient elimination of tumor cells. These molecules opened up new perspectives for anticancer therapy. Here, we will overview the structural features of the various types of antibody fragments and their applications for anticancer therapy as separate molecules and as part of complex conjugates or structures. Mechanisms of antitumor action of antibody fragments as well as their advantages and disadvantages for clinical application will be discussed in this review.

背景与目标： 单克隆抗体（mAbs）是一类重要的治疗药物，已批准用于治疗多种类型的恶性肿瘤。但是，在某些情况下，常规mAb的应用在抗癌免疫治疗中有一些局限性。这些局限性包括功效不足和副作用。抗体的抗原结合片段具有巨大的潜力，可以克服常规mAb的缺点，例如难以穿透实体瘤和Fc介导的旁观者激活免疫系统。抗体片段保留了抗原特异性和常规mAb的部分功能特性，同时具有更好的渗透性并大大降低了不良反应水平。抗体工程学中的最新优点允许产生具有改进的结构和性质的不同类型的抗体片段，以有效消除肿瘤细胞。这些分子为抗癌治疗开辟了新的前景。在这里，我们将概述各种类型的抗体片段的结构特征及其作为单独分子以及作为复杂缀合物或结构的一部分在抗癌治疗中的应用。在本综述中将讨论抗体片段的抗肿瘤作用机制以及它们在临床应用中的优缺点。

BACKGROUND & AIMS: :A Gram-staining-negative, rod-shaped, non-spore-forming bacterium isolated as a contaminant from a biopharmaceutical process (strain CCUG 53591(T)) was studied for its taxonomic allocation. On the basis of 16S rRNA gene sequence similarity data, this strain was clearly allocated to the genus Herminiimonas. Herminiimonas saxobsidens was shown to be the most closely related species on the basis of 16S rRNA gene sequence similarity (99.9 %), followed by Herminiimonas glaciei (99.6 %) and Herminiimonas arsenicoxydans (98.8 %). Strain ND5, previously reported as H. glaciei, but showing 100 % 16S rRNA gene sequence similarity to strain CCUG 53591(T), was included in the comparative study. Similarities to all other species of the genus Herminiimonas were below 98.0 %. Chemotaxonomic data (major ubiquinone, Q-8; major polar lipids, phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol; and major fatty acids, C(17 : 0) cyclo, C(19 : 0) cyclo ω8c and C(16 : 0,) with C(10 : 0) 3-OH as hydroxylated fatty acid) supported the affiliation of the isolate to the genus Herminiimonas. DNA-DNA hybridization results (mean values) for strain CCUG 53591(T) with H. saxobsidens CCUG 59860(T) (34 %), H. glaciei DSM 21140(T) (57 %), H. arsenicoxydans DSM 17148(T) (17 %) and Herminiimonas fonticola S-94(T) (11 %) clearly supported the separate taxonomic position of this strain. Strain ND5 showed DNA-DNA similarities of 78, 56 and 52 % to strain CCUG 53591(T), H. glaciei DSM 21140(T) and H. saxobsidens CCUG 59860(T), respectively. Phenotypic differentiation of the isolate from the most closely related species was possible by various features. Hence, strain CCUG 53591(T) represents a novel species, for which the name Herminiimonas contaminans sp. nov. is proposed, with the type strain CCUG 53591(T) ( = CCM 7991(T)). Strain ND5 is a second strain of this species.

背景与目标： ：从生物制药过程（菌株CCUG 53591（T））中分离出作为污染物的革兰氏染色阴性，杆状，无芽孢的细菌，对其分类进行了研究。根据16S rRNA基因序列相似性数据，该菌株被明确分配给Herminiimonas属。根据16S rRNA基因序列相似性，沙门氏菌是最密切相关的物种（99.9％），其次是冰河埃米菌（99.6％）和亚砷酸单胞菌（98.8％）。 ND5菌株以前被报道为H. glaciei，但显示出与CCUG 53591（T）菌株具有100 %%的16S rRNA基因序列相似性，被纳入比较研究。与Herminiimonas属的所有其他物种的相似性低于98.0％。化学分类数据（主要泛醌Q-8;主要极性脂质，磷脂酰乙醇胺，磷脂酰甘油和二磷脂酰甘油;以及主要脂肪酸C（17：0）环，C（19：0）环ω8c和C（16：0，） C（10：0）3-OH为羟基脂肪酸）支持分离物与Herminiimonas的隶属关系。菌株CCUG 53591（T）与萨克斯链球菌CCUG 59860（T）（34％），冰川嗜血杆菌DSM 21140（T）（57％），砷氧化单链菌DSM 17148（T）的DNA-DNA杂交结果（平均值） ）（17％）和Herminiimonas fonticola S-94（T）（11％）清楚地支持了该菌株的单独分类学位置。 ND5菌株与CCUG 53591（T），H。glaciei DSM 21140（T）和Saxobsidens CCUG 59860（T）的DNA-DNA相似性分别为78％，56％和52％。分离物从最密切相关的物种的表型分化可能有多种特征。因此，菌株CCUG 53591（T）代表了一个新物种，其名称为Herminiimonas contaminans sp。十一月提出了一种类型为CCUG 53591（T）（= CCM 7991（T））的传感器。菌株ND5是该菌种的第二个菌株。

BACKGROUND & AIMS: :Production of biopharmaceuticals from mammalian cells requires generation of master, working and post-production cell banks of high quality under GMP conditions. An optimal cryopreservation strategy is needed for each new production cell line, particularly with regard to establishing production processes that are completely devoid of serum or even any animal components and to ensuring robust thaw performance for reliable production. Here, we describe a novel strategy employing flow-cytometric (FC) analysis of Annexin V-stained cells for high-throughput characterization of cell banks. Our data show that this method enables predictive evaluation of a cryopreservation strategy as early as 6h after thawing of cells. Furthermore, a broad study is presented characterizing various factors that may influence the quality of serum-free production cell banks from NSO and CHO cell lines. These results demonstrate how FC-based analysis can be used for development of future state-of-the-art cryopreservation strategies.

背景与目标： ：从哺乳动物细胞生产生物药物需要在GMP条件下生成高质量的主，工作和后期生产细胞库。每个新的生产细胞系都需要一种最佳的冷冻保存策略，尤其是在建立完全不含血清或什至没有动物成分的生产过程以及确保可靠的解冻性能以实现可靠生产方面。在这里，我们描述了一种新的策略，利用膜联蛋白V染色的细胞的流式细胞术（FC）分析进行细胞库的高通量表征。我们的数据表明，这种方法能够在细胞解冻后的6小时内对冷冻保存策略进行预测性评估。此外，提出了一项广泛的研究，描述了可能影响来自NSO和CHO细胞系的无血清生产细胞库质量的各种因素。这些结果证明了如何将基于FC的分析用于未来最先进的冷冻保存策略的开发。