流行性感冒裂解疫苗的研制

作为换代产品,流行性感冒裂解疫苗的研制已取得突破性进展,根据WHO有关规程的规定和大量的试验研究结果。完整建立了该疫苗的生产工艺和生产质量控制系统,完成了“流行性感冒裂解疫苗制造及检定规程”等规定性文件的起草和审核工作,以中试规模连续生产了三批疫苗并全部自检合格,通过疫苗稳定性试验,效力试验,异常毒性试验及过敏性试验等观察。进一步肯定了疫苗的质量。     

重组双功能水蛭素质量标准研究

以生色底物法测定抗凝血酶活性,比浊法测定抗血小板聚集活性,还原型SDS-PAGE测定分子量,SDS-PAGE和反相高效液相色谱测定纯度,毛细管电泳法测定等电点,胰蛋白酶酶切后进行肽图分析,其余检测项目按《中国药典》2005版三部规定进行。结果显示,用建立的方法对原液和成品进行了检定,各项指标均符合《人用重组DNA制品质量控制技术指导原则》和《中国药典》2005版三部的要求。建立的质控方法和质量标准具有保证产品安全、有效、质量可控的特点,可用于重组双功能水蛭素产品的常规检定。     

Supplementary quality controls of plasma fractions for manufactures and consumers

As a consequence of the increasing use of plasma fractions, there is an ever-growing abundance of such preparations. These preparations may be divided into three main groups: clotting factors, immunoglobulins, and plasma protein solutions. Frequency of component-induced side-effects are inversely proportional to those components' quality. Pharmacopoeas presently require only the following quality criteria for most plasma preparations: declaration of sort and concentration of the active component and stabilizers, sterility, and the absence of pyrogens. Of great importance is the origin of the raw material and the methods used to harvest the plasma, as well as the molecular condition of the proteins. While we can assume that presently-available preparations meet minimum requirements, it can be demonstrated that similar preparations can in no case be considered equivalent when immunoelectrophoresis, ultracentrifugation and gel filtration are used as additional quality control tests. Especially immunoelectrophoresis and gel filtration are tests easily available to the clinician to help him detect and thus select the optical product.     

Expert examination of the immunogenic activity and toxicity of the pertussis component of adsorbed and non-adsorbed diphtheria-tetanus-pertussis vaccines. Results of interlaboratory investigations under the WHO project "Reactogenicity and Toxicity of DPT Vaccines"

Since the DPT vaccine is broadly used for the prevention of diphtheria, tetanus and pertussis' its preparations should meet special quality requirements. The present study was aimed at evaluating the reproducibility of laboratory methods utilized to assess the protective activity and toxicity of the pertussis component as well as at examining the feasibility of expert estimations of product quality to enhance the validity of findings. The results of interlaboratory comparative examination of quality parameters of the tested preparations revealed that the routine laboratory quality control methods were not sufficiently standardized as their application in different laboratories did not always produce identical results. The WHO criteria to evaluate the toxicity of pertussis vaccines are far from perfect since vaccines with pronounced toxicity can only be distinguished from those with moderate to mild toxicity after the administration to tested mice of vaccines at a single infant dose of 0.5 ml. To enhance the reproducibility of methods employed in the laboratory, appropriate standard specimens of the preparation should be used serving as a measure of different quality parameters; all test conditions should be also standardized as far as possible. To enable objective quality evaluation of medical biological preparations, the degree of experiment reproducibility should be regularly verified in interlaboratory tests on an international scale as well as inside those countries which have several manufacturers of a given preparation. It appears expedient to set up an international system of expert evaluation of quality of biological preparations by appointing several regional and national centres which meet the requirements for expert laboratories.     

人降钙素基因相关肽脂质体质量标准的研究

建立完整的人降钙素基因相关肽脂质体(LipohCGRP)药物的质量标准。用家兔球睫膜血管扩张法测定hCGRP的生物学活性;采用RPHPLC、等电聚焦、薄层层析等方法分别测定样品的纯度、等电点和迁移率及脂质体嵌入率的测定,并按照中国生物制品规程的要求完成了甲醇和氯仿等残留物质的分析和动物安全试验。建立了活性测定方法,能准确测定样品中不到1ng的样品活性单位,连续三批样品的各项指标均符合质量标准的要求。建立了脂质体多肽药物的质量标准,并用于质量检定。     

A Comparison of Methods to Measure Operator Protection Factors in Open-fronted Microbiological Safety Cabinets

Comparative tests to measure operator protection factors in microbiological safety cabinets in accordance with British Standard 5726 have demonstrated good agreement in the results obtained by a microbiological method using a Collison nebulizer and the technique producing an aerosol of potassium iodide. Either method is suitable for testing for operator protection factors in Class I and Class II safety cabinets.
The Collison nebulizer should be considered as the standard aerosol generator for the microbiological method; alternative nebulizers meeting the general requirements of BS 5726 should be compared in performance with this nebulizer if they are to be used for containment tests.
Any microbiological safety cabinet specified for a new installation should have been 'type' tested to ensure compliance with BS 5726. However, in order to ensure adequate performance, on-site commissioning tests (and routine maintenance checks thereafter) are necessary to verify that air velocity, filtration and operator protection factor requirements are met.     

Identification of nonlinear models of biological membranes using the voltage-clamp method

The method for identification of nonlinear systems proposed in 1952 by Hodgkin and Huxley is mathematically justified. A procedure for the application of this method is developed, including the development of the structure of a mathematical model, carrying out a series of tests with special chosen signals, and determination of unknown parameters. Basic requirements for the admissible sets of input and output signals and to the system operator have been determined. It is shown that this operator should be totally continuous and that the minimum number of unknown parameters and the minimum complexity of the operator structure should give an approximation of the necessary quality. The pros and cons of the Hodgkin-Huxley and Noble mathematical models and the methods used for their development are discussed. A structure for the operator for the identification of mathematical models of excitable membranes with a large number of membrane currents is proposed. It is found that the nonlinear electrical properties of biological membranes can be identified using tests with other types of “clamped” parameters, such as the current, ramp voltage, etc.     

Guidelines for safety tests and registration of bacterial pesticides

Guidelines have been formulated for safety tests and registration of new microbial pesticides containing pathogenic or inhibitory bacteria. These guidelines are intended for the advice of interested firms and governmental agencies. They are based on over a decade's experience and intense recent activity in this field. For the registration of a new microbial product, information is required on the identity of the new bacterium, its biological properties, production, formulation, quality control, application and efficacy. For safety assessment a series of tests on the infectivity, toxicity and allergenicity etc. in laboratory mammals is arranged in 3 tiers. Entirely negative results in tier 1 indicate acceptance of the product as safe. Positive results may lead to its rejection or to further lengthier tests in tiers 2 and 3, culminating in rejection or acceptance after a risk-benefit analysis, possibly with restrictions concerning its use required on the label. Data on residues, inactivation and degradability may be necessary. Information on exposure of humans during production and use are particularly valuable. The intended use of the product greatly influences both safety test requirements and tests on non-target organisms and wildlife, e.g. bees, important predators and parasites of the target species, earthworms, birds and fish. Also phytotoxicity and phytopathogenicity must be considered. Aspects of current interest are critically discussed. Any fundamental studies not justified as mandatory requirements for testing a new product should be funded by non-industrial sources.     

Microbes in food processing technology

Abstract: There is an increasing understanding that the microbial quality of a certain food is the result of a chain of events. It is clear that the microbial safety of food can only be guaranteed when the overall processing, including the production of raw materials, distribution and handling by the consumer are taken into consideration. Therefore, the microbiological quality assurance of foods is not only a matter of control, but also of a careful design of the total process chain. Food industry has now generally adapted quality assurance systems and is implementing the Hazard Analysis Critical Control Point (HACCP) concept. Rapid microbiological monitoring systems should be used in these cases. There is a need for rapid and simple microbiological tests which can be adapted to the technology and logistics of specific production processes. Traditional microbiological methods generally do not meet these high requirements. This paper discusses the tests, based on molecular biological principles, to detect and identify microbes in food-processing chains. Tests based on DNA technology are discussed, including in vitro DNA amplification like the polymerase chain reaction (PCR) method and identifications based on RFLP, RAPD and DNA fingerprinting analysis. PCR-haled methodology can be used for the rapid detection of microbes in food manufacturing environments. In addition, DNA fingerprinting methods are suitable for investigating sources and routes of microbial contamination in the food cycle.     

乙型肝炎诊断试剂国家质控标准的建立

按新的方案建立了用于乙型肝炎诊断试剂的国家质控标准。包括用WHO国际标准品标化建立的我国HBsAg、抗-HBs定量标准品,对试剂的特异性、灵敏度、精密性、线性进行质量控制检定的系列参比品,及经过实验制订的检定标准。经近年来的实际应用,对国产试剂质量的提高起到了推动作用。     

The data quality foundation in OMNIITOX information system

Extended Summary Goal and Scope  This paper describes the data quality foundation for the OMNIITOX information system, with regard to data quality requirements and maintenance. Main Features  The data quality concept applied in the OMNIITOX project is based on results that are successfully used in other contexts (Fig. 1). Specific data quality requirements have been developed, which are applied in data acquisition and data review. The data quality requirements and quality maintenance is supported by the OMNIITOX concept model, which supplies a common language for the users from the different disciplines. Results and Discussion  The quality requirements and the concept model have been tested during data acquisition for the information system, and have been found to be a successful framework. The results from practical work demonstrate that it is possible to have a common understanding of the concept model and quality requirements. The user's experience the initial phase of learning the concepts and quality requirements as tedious. Nevertheless, experiences show that the users will benefit from the education during the further work. A considerable amount of data has been acquired based on the data quality foundation. Conclusions  The data quality foundation has been a successful framework for managing data quality in the design and establishment of the OMNIITOX information system. The data quality concept, the concept model, and specific quality requirements are applied as quality management tools to find, acquire, document, and interpret the information. Several quality issues deriving from interdisciplinary differences between the different users have been resolved, which facilitates a common understanding and reduces the risk of misinterpretations and misuse of information. Recommendations and Outlook  An important task for the finalization of the OMNIITOX project is to secure the credibility and long-term stability of the information system. The OMNIITOX information system will somewhat have less potential to support the toxicology related assessments in Europe if any of the involved disciplines chooses to reject or not to apply the concept model and the quality requirements.     

Critical testing and parameters for consideration when manufacturing and evaluating tumor–associated antigen-specific T cells

The past year has seen remarkable translation of cellular and gene therapies, with U.S. Food and Drug Administration (FDA) approval of three chimeric antigen receptor (CAR) T-cell products, multiple gene therapy products, and the initiation of countless other pivotal clinical trials. What makes these new drugs most remarkable is their path to commercialization: they have unique requirements compared with traditional pharmaceutical drugs and require different potency assays, critical quality attributes and parameters, pharmacological and toxicological data, and in vivo efficacy testing. What's more, each biologic requires its own unique set of tests and parameters. Here we describe the unique tests associated with ex vivo–expanded tumor-associated antigen T cells (TAA-T). These tests include functional assays to determine potency, specificity, and identity; tests for pathogenic contaminants, such as bacteria and fungus as well as other contaminants such as Mycoplasma and endotoxin; tests for product characterization, tests to evaluate T-cell persistence and product efficacy; and finally, recommendations for critical quality attributes and parameters associated with the expansion of TAA-Ts.     

Single‐step affinity purification of enzyme biotherapeutics: A platform methodology for accelerated process development

Downstream sample purification for quality attribute analysis is a significant bottleneck in process development for non‐antibody biologics. Multi‐step chromatography process train purifications are typically required prior to many critical analytical tests. This prerequisite leads to limited throughput, long lead times to obtain purified product, and significant resource requirements. In this work, immunoaffinity purification technology has been leveraged to achieve single‐step affinity purification of two different enzyme biotherapeutics (Fabrazyme® [agalsidase beta] and Enzyme 2) with polyclonal and monoclonal antibodies, respectively, as ligands. Target molecules were rapidly isolated from cell culture harvest in sufficient purity to enable analysis of critical quality attributes (CQAs). Most importantly, this is the first study that demonstrates the application of predictive analytics techniques to predict critical quality attributes of a commercial biologic. The data obtained using the affinity columns were used to generate appropriate models to predict quality attributes that would be obtained after traditional multi‐step purification trains. These models empower process development decision‐making with drug substance‐equivalent product quality information without generation of actual drug substance. Optimization was performed to ensure maximum target recovery and minimal target protein degradation. The methodologies developed for Fabrazyme were successfully reapplied for Enzyme 2, indicating platform opportunities. The impact of the technology is significant, including reductions in time and personnel requirements, rapid product purification, and substantially increased throughput. Applications are discussed, including upstream and downstream process development support to achieve the principles of Quality by Design (QbD) as well as integration with bioprocesses as a process analytical technology (PAT). © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:708–717, 2014     

一种简便易行核酸疫苗质粒纯化工艺的开发

介绍了一种成本低、步骤少、简单易行的质粒纯化制检工艺。该工艺选择优势产生超螺旋质粒的大肠杆菌菌株以无蛋白质培养基进行发酵罐培养,采用碱裂解法,对质粒制备过程中所用的层析吸附材料、核酸结合溶液、去除内毒素等杂质的方法和浓缩等步骤进行了实用性改进,并建立了相应的检定方法,所得质粒的纯度达到临床级要求。     

BSCC Code of Practice – fine needle aspiration cytology

The British Society for Clinical Cytology Code of Practice on fine needle aspiration cytology complements that on exfoliative cytopathology, which was published in the last issue ( Cytopathology 2009; 20 :211–23). Both have been prepared with wide consultation within and outside the BSCC and have been endorsed by the Royal College of Pathologists. A separate code of practice for gynaecological cytopathology is in preparation. Fine needle aspiration (FNA) cytology is an accepted first line investigation for mass lesions, which may be targeted by palpation or a variety of imaging methods. Although FNA cytology has been shown to be a cost-effective, reliable technique its accurate interpretation depends on obtaining adequately cellular samples prepared to a high standard. Its accuracy and cost-effectiveness can be seriously compromised by inadequate samples. Although cytopathologists, radiologists, nurses or clinicians may take FNAs, they must be adequately trained, experienced and subject to regular audit. The best results are obtained when a pathologist or an experienced and trained biomedical scientist (cytotechnologist) provides immediate on-site assessment of sample adequacy whether or not the FNA requires image-guidance. This COP provides evidence-based recommendations for setting up FNA services, managing the patients, taking the samples, preparing the slides, collecting material for ancillary tests, providing rapid on-site assessment, classifying the diagnosis and providing a final report. Costs, cost-effectiveness and rare complications are taken into account as well as the time and resources required for quality control, audit and correlation of cytology with histology and outcome. Laboratories are expected to have an effective quality management system conforming to the requirements of a recognised accreditation scheme such as Clinical Pathology Accreditation (UK) Ltd.     

Data pre-processing in liquid chromatography-mass spectrometry-based proteomics

MOTIVATION: In a liquid chromatography-mass spectrometry (LC-MS)-based expressional proteomics, multiple samples from different groups are analyzed in parallel. It is necessary to develop a data mining system to perform peak quantification, peak alignment and data quality assurance. RESULTS: We have developed an algorithm for spectrum deconvolution. A two-step alignment algorithm is proposed for recognizing peaks generated by the same peptide but detected in different samples. The quality of LC-MS data is evaluated using statistical tests and alignment quality tests. AVAILABILITY: Xalign software is available upon request from the author.     

The BSCC Code of Practice – exfoliative cytopathology (excluding gynaecological cytopathology)

Exfoliative cytopathology (often referred to as non‐gynaecological cytology) is an important part of the workload of all diagnostic pathology departments. It clearly has a role in the diagnosis of neoplastic disease but its role in establishing non‐neoplastic diagnoses should also be recognised. Ancillary tests may be required to establish a definitive diagnosis. Clinical and scientific teamwork is essential to establish an effective cytology service and staffing levels should be sufficient to support preparation, prescreening, on‐site adequacy assessment and reporting of samples as appropriate. Routine clinical audit and histology/cytology correlation should be in place as quality control of a cytology service. Cytology staff should be involved in multidisciplinary meetings and appropriate professional networks. Laboratories should have an effective quality management system conforming to the requirements of a recognised accreditation scheme such as Clinical Pathology Accreditation (UK) Ltd. Consultant pathologists should sign out the majority of exfoliative cytology cases. Where specimens are reported by experienced biomedical scientists (BMS), referred to as cytotechnologists outside the UK, this must only be when adequate training has been given and be defined in agreed written local protocols. An educational basis for formalising the role of the BMS in exfoliative cytopathology is provided by the Diploma of Expert Practice in Non‐gynaecological Cytology offered by the Institute of Biomedical Science (IBMS). The reliability of cytological diagnoses is dependent on the quality of the specimen provided and the quality of the preparations produced. The laboratory should provide feedback and written guidance on specimen procurement. Specimen processing should be by appropriately trained, competent staff with appropriate quality control. Microscopic examination of preparations by BMS should be encouraged wherever possible. Specific guidance is provided on the clinical role, specimen procurement, preparation and suitable staining techniques for urine, sputum, semen, serous cavity effusion, cerebrospinal fluid, synovial fluid, cyst aspirates, endoscopic specimens, and skin and mucosal scrapes.     

Strategies for computer modeling

Modeling is a ubiquitous and often misunderstood enterprise in which data from diverse disciplines are analyzed by techniques from other diverse disciplines in an attempt to confirm or falsify a set of hypotheses about the real world. Guidelines are offered for designing models to match the goals of modeling biological systems. Techniques for the construction and interpretation of models are discussed. The requirements for credibility of models are detailed, and tests are suggested for their validation.     

Statistical tests as inappropriate tools for data analysis performed on non-random samples of plant communities

Many authors apply statistical tests to sets of relevés obtained using non-random methods to investigate phytosociological and ecological relationships. Frequently applied tests include thet-test, ANOVA, Mann-Whitney test, Kruskal-Wallis test, chi-square test (of independence, goodness-of-fit, and homogeneity), Kolmogorov-Smirnov test, concentration analysis, tests of linear correlation and Spearman rank correlation coefficient, computer intensive methods (such as randomization and re-sampling) and others. I examined the extent of reliability of the results of such tests applied to non-random data by examining the tests requirements according to statistical theory. I conclude that when used for such data, the statistical tests do not provide reliable support for the inferences made because non-randomness of samples violated the demand for observations to be independent, and different parts of the investigated communities did not have equal chance to be represented in the sample. Additional requirements, e.g. of normality and homoscedasticity, were also neglected in several cases. The importance of data satisfying the basic requirements set by statistical tests is stressed.