scholarly journals Identification of lumbrokinase by fibrin-plate method, fibrin-zymography and liquid chromatography mass spectrometry

2019 ◽  
Author(s):  
Bingbing Ke’ ◽  
Ruiqing Xian ◽  
Hong Jiang ◽  
Jianghong Guo
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Eisenia Fetida ◽  
Plasminogen Activators ◽  
Liquid Chromatography Mass Spectrometry ◽  
Plate Method ◽  
Bovine Trypsin ◽  
Fibrin Plate ◽  
Human Plasminogen ◽  
Protein Components

AbstractLumbrokinase, extracted from the cultured earthworm of Eisenia fetida, has been widely used as biochemical medicine in China to prevent or treat thrombosis. In the present study, the mechanism of lumbrokinase was investigated using the fibrin plate method. The results revealed that lumbrokinase contained both fibrinolytic and kinase components. The method of fibrin-zymography was used to show the existence and activity of lumbrokinase, and we proved that the fibrin-zymogram gel could be adopted as the identification method of earthworm. Subsequently, the components were identified by mass spectrometry. According to the results, fibrinolytic related components existed in the drug. These proteins were further compared with other serine proteins. The result showed that the identified proteins were similar to human trypsin and bovine trypsin. Besides, some also exhibited similar characteristics with human plasminogen activators. The mentioned results demonstrated that lumbrokinase products contained two major groups of protein components, suggesting two different functions.

A Laconic Review on Liquid Chromatography Mass Spectrometry (LC-MS) Based Proteomics Technology in Drug Discovery

2021 ◽  
pp. 194-201
Author(s):  
Shivani Sharma ◽  
Navdeep Singh ◽  
Amar Deep Ankalgi ◽  
Arti Rana ◽  
Mahendra Singh Ashawat
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
New Product Development ◽  
Pharmaceutical Products ◽  
Liquid Chromatography Mass Spectrometry ◽  
Intact Proteins ◽  
Chromatography Mass Spectrometry ◽  
The Future ◽  
Protein Components ◽  
The Stability

Liquid chromatography mass spectrometry is a powerful technique which is used for the new product development, manufacturing, and to the control the stability or drug abuse. These techniques were commonly preferred in pharmacokinetics studies in pharmaceutical products. The principle involved in the LC-MS technique was justified in our study to understand the basic fundamentals of LC-MS. Also, it is used in combination with HPLC for chemical analysis. Many applications and the future prospects have been highlighted regarding the use for LC-MS in analytical chemistry. It is basically depends on the metabolites which are collected in this technique, furthermore the chemicals are analyzed according to the nature of suitability of LC-MS. This technique was helpful in the analysis of protein components identified in terms of pharmacovigilance, organic/inorganic hybrid nanoflowering. Also, our study highlights the techniques involved in proteomics. In LC-MS-based proteomics, complex mixtures of proteins are first subjected to enzymatic cleavage, then the resulting peptide products are analyzed using a mass spectrometer; this is in contrast to "top-down" proteomics, which deals with intact proteins and is limited to single protein mixtures. So, this review aims to highlight the basic introduction, and principle involved in liquid chromatography-mass spectrometry (LC-MS). Also, the advantages, or application of LC-MS were studied. Most importantly the LC-MS based proteomics, and the future aspects of LC-MS technology were studied in this review.

Evaluation of the Fibrin Plate Method for Estimating Plasminogen Activators

1972 ◽  
Vol 28 (01) ◽  
pp. 075-088 ◽  
Author(s):  
N. A Marsh ◽  
C. L Arocha-Pinango
Keyword(s):  
Tranexamic Acid ◽  
Plasminogen Activators ◽  
Heated Plate ◽  
Plasminogen Activation ◽  
Response Curves ◽  
Plate Method ◽  
Single Plate ◽  
Effect Of Additives ◽  
Threefold Increase ◽  
Fibrin Plate

SummaryA study was carried out in order to evaluate the Astrup and Mullertz fibrin plate method for estimating plasminogen activators.Choice of a suitable fibrinogen substrate was found to be the most important factor in setting up a workable assay. Many preparations contained a large proportion of non-clottable protein and plates made from these fibrinogens were usually unreliable. In addition, plasminogen content varied appreciably between preparations so that sensitivity of the method required careful calibration with each new batch of fibrinogen.The effect of additives in the fibrin plate was considered and it was found that calcium chloride alone was sufficient to ensure a stabilised plate which could be stored at 4° C for some time. The addition of tranexamic acid (AMCHA) was found to be a slightly more convenient way of estimating direct proteolytic activity, compared with the traditional heated plate. However neither method distinguished completely between proteolysis and plasminogen activation.In order to improve the precision of the method, the use of an analysis of variance technique has been studied. This technique provides information on the dose-response curves of test and unknown substances, and in addition produces an approximately threefold increase in precision over single plate tests.

Erratum: Recommendations for the Use of Liquid Chromatography-Mass Spectrometry in the Clinical Laboratory: Part I. Implementation and Management

2020 ◽  
Vol 10 (2) ◽  
pp. 179
Author(s):  
Kyunghoon Lee ◽  
Soo Young Moon ◽  
Serim Kim ◽  
Hyun-Jung Choi ◽  
Sang-Guk Lee ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Clinical Laboratory ◽  
Liquid Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry
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