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@#Objective To verify the applicability of Lowry2 method[the second method of Folin-phenol method of Chinese Pharmacopoeia(VolumeⅢ,2020 edition)]for the determination of protein content of intermediate products of component pertussis vaccine.Methods The standard and sample were precipitated and pretreated with deoxycholate and trichloroacetic acid to remove impurities,and then reacted with Folin-phenol,and the absorbance was measured at 750 nm wavelength.The standard curve was drawn with the concentration and absorbance of the standard,and the sample concentration was calculated. The method was verified for the specificity,linearity,accuracy and precision,and used to detect the protein content of intermediate products of three batches of component pertussis vaccine.Results There was no significant difference between the protein content in background buffer of intermediate product detected by this method and that in ultrapure water(t = 0. 277～1. 178,P = 0. 304～0. 795);The linear relationship of standard curve was good at the protein concentration of 0～50 μg/mL,each R~2> 0. 98;The spike recovery of accuracy verification was in the range of 90%～110%;The RSDs of repeatability and intermediate precision verification were both less than 3%. The detection results of protein content in the intermediate products of three batches of component pertussis vaccine all met the requirements of internal manufacturing and verification regulations.Conclusion Lowry2 method has good specificity,accuracy and precision,and can be used to determine the protein content of component pertussis intermediate products,which provides a basis for the applicability of Lowry2 method.
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Proteomics represented vital applications of technologies in the identification and quantification of high to moderate proteins (cellular signalling networks) found in biological matrix such as tissues, cells and fluids. Proteomics based technical knowledge is applied and verified in several preclinical research settings such as invention of diagnostic markers for specific disease and have shown to be increased in clinical applications. Extensive studies on proteomics resulted in detection of biomarkers that have been highly advanced in using diseases for cancer, lungs, cardiovascular, renal and neuro-regenerative and Parkinson's disease by introducing human origins for biocompatibility such as urine and serum. Advancement in the proteomic methods is conferring candidate right direction for clinical usage. In this review, recent developments and widely used proteomics approaches such as Mass Spectrometry (MS), Microarray chips are elaborately addressed and also focused merits and demerits of commonly used advanced approaches such as Selected Reaction Monitoring (SRM), Parallel Reaction Monitoring (PRM) and Data Independent Acquisition (DIA) and other used proteomics and that roles, in order to aid clinicians, were also discussed in the light of biomedical applications.
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Objective To observe the clinical therapeutic effect of Bushen Huoxue formula for treatment of diabetic nephropathy at G3a stage. Methods Sixty patients with stage G3a diabetic nephropathy were admitted to the Department of Nephrology of Affiliated Hospital of Tianjin Institute of Traditional Chinese Medicine from June 2017 to June 2018, and according to difference in treatment, they were divided into an integrated traditional Chinese and western medicine treatment group and a western medicine treatment group with 30 cases in each group. The two groups were treated with Huangkui capsule 2.5 g/time, 3 times a day; the integrated traditional Chinese and western medicine treatment group was additionally given Bushen Huoxue formula, one dose daily, twice a day taken orally; 2 months for 1 course. The changes of 24-hour urinary protein, serum creatinine (SCr) and urea nitrogen (BUN) were observed after 3 courses of treatment. Results After 3 courses of treatment, the levels of 24-hour urinary protein, SCr and BUN of the integrated traditional Chinese and western medicine treatment group were lower than those of the western medicine treatment group [24-hour urinary protein quantification (g): 1.45±0.26 vs. 2.11±0.35, SCr (μmol/L): 105.15±12.31 vs. 158.32±17.26, BUN (mmol/L): 7.26±2.41 vs. 12.87±3.24], the differences being statistically significant (all P < 0.05). The total effective rate of integrated traditional Chinese and western medicine treatment group was significantly higher than that of western medicine treatment group [86.67% (26/30) vs. 53.33% (16/30), P < 0.05]. Conclusion Bushen Huoxue formula in the treatment of patients with diabetic nephropathy at stage G3a can decrease their urinary protein and SCr, BUN significantly.
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Introducción: La Inmunodifusión radial simple es una técnica con fundamento inmunológico confiable por su especificidad para la cuantificación de inmunoglobulinas principales y se emplea también para otras proteínas. Las placas de Inmunodifusión comerciales se ofertan con un número determinado de pocillos donde se coloca la muestra biológica que contiene la proteína a cuantificar. Objetivo: Evaluar la sensibilidad y la especificidad de la modificación introducida para optimizar el uso de las placas de inmunodifusión radial simple de la marca SIEMENS por aumento del número de muestras por placas. Material y Métodos: Se presenta una innovación que permite optimizar el área biológicamente activa de la placa no utilizada para emplearla para la cuantificación de otras muestras. Se realizan montajes paralelos de muestras de controles en los pocillos tradicionales y en los realizados en los espacios disponibles para cuantificar IgG y albúmina para suero y líquido cefalorraquídeo. Resultados: La sensibilidad del empleo por el método tradicional y por el nuevo no presenta diferencias significativas. En cuanto a la especificidad tampoco existen diferencias significativas menos en las placas para cuantificar albúmina en suero por lo que se recomienda diluir la muestra de suero antes de ser utilizada en el área disponible. En el caso de las placas NOR y LC Partigen® el número de muestras a ser beneficiadas con la cuantificación se duplica, pero de igual manera puede ser aplicada en otras placas de otras firmas comerciales. Conclusiones: Esta innovación permite hacer un uso óptimo de las placas de inmunodifusión con el consiguiente ahorro de material de importación y se puede aplicar fácilmente en todos los laboratorios del país(AU)
Introduction: Single radial immunodiffusion assay is a technique with immunological base, which is reliable because of its specificity in the quantification of main immunoglobulins, although it is also used for other proteins. Commercial immunodiffusion plates are offered with a determined number of holes where the biological samples containing protein to be quantified are placed. Objective: To evaluate the sensitivity and specificity of the modification implemented to optimize the usage of single radial immunodiffusion plates from Siemens by increasing the number of samples in the plates. Materials and Methods: An innovating procedure that allows to optimize the non-used biologically active area and use it in the quantification of other samples is presented. A parallel quantification of control samples from traditional holes and the other ones opened in available spaces was performed in order to quantify IgG and albumin in serum and in cerebrospinal fluid. Results: Sensitivity was not affected significantly between the normal plates and the usage of the new procedure. Regarding specificity, there are also no significant differences except in the plates used to quantify serum albumin; so, it is recommended to dilute serum samples before the application. In case of NOR and LC Partigens®, this proposed modification duplicates the number of samples to be quantified in each plate, but otherwise, it could be applied in other commercial immunoplates. Conclusions: This innovation allows to make an optimal usage of immunodiffusion plates with the consequent saving of import materials, which can be easily applied in all the laboratories of the country(AU)
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Humans , Laboratory Equipment , Immunodiffusion/methods , Mandatory TestingABSTRACT
@#Quantitative proteomics is a mass spectrometry-based toolkit used to analyze and quantify entire proteins contained in whole cells, tissues or organisms. It has become an increasingly important element in exploring the mechanism of various biological processes such as discovering novel biomarkers and unknown drug targets. Emerging advances in biological mass spectrometry instrumentation and data acquisition methodologies have provided a state-of-the-art platform for protein quantification, prompting the research of proteomics evolving from the simple qualitative to the accurate quantitative approach. This review aims to introduce the most recent advancements in mass spectrometry instrumentation and methodologies of data acquisition, focusing on their characteristics and applying fields. It also highlights several significant applications of biological mass spectrometry in pharmaceutical research such as quantifitation of drug transporters and metabolizing enzymes, and pharmacokinetic study of therapeutic peptides and proteins.