The Adverse Effect of Glycation of Human Serum Albumin on its Preservative Activity in the Freeze-Drying and Accelerated Degradation of Alkaline Phosphatase

Author:

Ford Annette W1,Allahiary Zahra1

Affiliation:

1. Standards Processing Division, National Institute for Biological Standards and Control, Blanche Lane, South Minims, Potters Bar, Hertfordshire EN6 3QG, UK

Abstract

Abstract The effects of a glycated protein additive on the stability of freeze-dried biological standards were studied using alkaline phosphatase as a model. Alkaline phosphatase was formulated with artificially glycated albumin, freeze-dried, sealed into glass ampoules and subjected to accelerated degradation studies at temperatures from —20 to 56°C. Alkaline phosphatase, freeze-dried without an additive in neutral buffer, lost over 95% of its activity, but when freeze-dried with human serum albumin it retained approximately 70% of the initial activity. Both deliberately glycated and native albumin protected approximately 70% of the initial activity on freeze-drying and this protection was maintained during storage of the freeze-dried product at temperatures of 20°C or below for up to 16 weeks. At 37°C or above, alkaline phosphatase activity was lost in a time- and temperature-related manner with changes appearing in the SDS-PAGE gels and FPLC chromatograms but, with the artificially glycated albumin formulations, this loss of activity and the changes in the gels and chromatograms happened earlier and at lower temperatures. Formulations with trehalose at 1% w/v and 15% w/v, but without albumin, preserved some 40% of alkaline phosphatase activity following freeze-drying. Further, approximately half of that activity was maintained after 16 weeks' storage at all temperatures up to 56°C by the 15% trehalose without albumin. This study indicates that, in the formulation of freeze-dried biological standards, if albumin is to be used, the initial degree of glycation should be kept to a minimum and combinations of albumin and reducing sugar should be avoided. Trehalose 15% provides an acceptable alternative where exposure to high temperature is likely to occur.

Publisher

Oxford University Press (OUP)

Subject

Pharmaceutical Science,Pharmacology

Reference28 articles.

1. Pharmacological modulation of the advanced glycosylation reaction;Brownlee,1989

2. Additives to biological substances IV. The stability of lactose as a carrier for biological standards;Calam;J. Biol. Stand.,1988

3. International biological standards and reference preparations;Campbell;J. Biol. Stand.,1974

4. Stabilization of dry phospholipid bilayers and proteins by sugars;Crowe;Biochem. J.,1987

5. Disc electrophoresis II. Method and application to human serum proteins;Davis;Ann NY Acad. Sci.,1964

Cited by 5 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3