Long term investigation of formulation buffers to mitigate stability issues of conjugated critical reagents

IF 1.6 4区 医学 Q4 BIOCHEMICAL RESEARCH METHODS
Glenn T. Miller , Teresa M. Caiazzo , Alison Joyce
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引用次数: 0

Abstract

Stability of conjugated critical reagents supporting ligand binding assays to enable biotherapeutic drug development is a universal concern. Formulation buffer employed for long-term cold storage may be key to mitigate protein aggregation issues. We investigated biophysical and functional attributes of murine mAb and human multispecific drug labeled with biotin, ruthenium, and Alexa fluor 647 frozen at −80 °C in PBS or a protein storage buffer for 3–15 months. Aggregation was observed at 4 months in mAb A-Ru (11.2%) and -Alexa (10%) in PBS followed by precipitation and reduced biological binding at 15 months. Increased aggregation in drug Ru (11.7%, 6 months) and Alexa (6.9%, 15 months) were noted but without impact on performance. There were no observations with biotin labeled reagents.

长期研究配方缓冲液,以缓解共轭关键试剂的稳定性问题。
支持配体结合检测以促进生物治疗药物开发的共轭关键试剂的稳定性是一个普遍关注的问题。用于长期冷藏的制剂缓冲液可能是缓解蛋白质聚集问题的关键。我们研究了用生物素、钌和 Alexa fluor 647 标记的小鼠 mAb 和人类多特异性药物的生物物理和功能属性,这些药物在 -80 °C 的 PBS 或蛋白质储存缓冲液中冷冻 3-15 个月。在 PBS 中的 mAb A-Ru(11.2%)和 -Alexa(10%)在 4 个月时出现聚集,15 个月时出现沉淀和生物结合力降低。药物 Ru(11.7%,6 个月)和 Alexa(6.9%,15 个月)中的聚集现象有所增加,但对性能没有影响。生物素标记的试剂则没有出现这种情况。
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来源期刊
CiteScore
4.10
自引率
0.00%
发文量
120
审稿时长
3 months
期刊介绍: The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells. In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.
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