CN110895282A - 时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法 - Google Patents

时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法 Download PDF

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CN110895282A
CN110895282A CN201811059144.9A CN201811059144A CN110895282A CN 110895282 A CN110895282 A CN 110895282A CN 201811059144 A CN201811059144 A CN 201811059144A CN 110895282 A CN110895282 A CN 110895282A
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labeling reagent
formula
antibody labeling
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汪劲能
周智慧
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Wuxi Jing Jing Biological Technology Co Ltd
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex

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Abstract

本发明涉及一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法，抗体标记试剂配方为磷酸氢二钠3%~6%，磷酸二氢钠2%~5%，2‑吗啉乙磺酸（MES）8%~15%，1‑(3‑二甲氨基丙基)‑3‑乙基碳二亚胺（EDC）40%~60%，N‑羟基琥珀酰亚胺（NHS）15%~20%，总质量组成为100%。本发明提供的抗体标记试剂配方不但提高了偶联效率，还保证了荧光微球与抗体偶联物的活性和稳定性，降低了生产成本。

Description

时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法
技术领域
本发明涉及一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法。
背景技术
蛋白质偶联是将小分子物质（如药物、抗原）或大分子物质（如酶、蛋白毒素等）以共价键的方式连接于蛋白质分子，以制备人工抗原、酶标抗体、免疫毒素等。蛋白质偶联技术在生物学和医学领域得到越来越广泛的应用。免疫荧光技术是将免疫学方法(抗原抗体特异结合)与荧光标记技术结合起来研究特异蛋白抗原在细胞内分布的方法。由于荧光素所发的荧光可在荧光显微镜下检出，从而可对抗原进行细胞定位。该技术的主要特点特异性强、敏感性高、速度快，但淬灭率高，不稳定，不易保存。
常用的偶联方法如戊二醛法、碳二亚胺法、过碘酸钠氧化法等不可避免地会产生酶或抗体的自身交联产物或多聚物，致使偶联效率降低、结合物活性减弱。目前抗体的偶联技术相对不是很成熟，偶联过程中抗体的偶联效率比较低。
发明内容
针对上述，本发明提供一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法，不但提高了偶联效率，还保证了荧光微球与抗体偶联物的活性和稳定性，降低了生产成本。
本发明所用的技术方案是，抗体标记试剂配方包括磷酸氢二钠，磷酸二氢钠，2-吗啉乙磺酸（MES），1-(3-二甲氨基丙基)-3-乙基碳二亚胺（EDC），N-羟基琥珀酰亚胺（NHS）。其中，磷酸氢二钠3%~6%，磷酸二氢钠2%~5%，2-吗啉乙磺酸（MES）8%~20%，1-(3-二甲氨基丙基)-3-乙基碳二亚胺（EDC）40%~60%，N-羟基琥珀酰亚胺（NHS）15%~30%，总质量组成为100%。
进一步，MES溶液中加入EDC和NHS，用磷酸钠缓冲液调节pH至7.2-9.0。
进一步，配制的MES溶液pH=4.6-6.5。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅用以解释本发明，并不用于限定本发明。
下面结合实施例对本发明的应用原理作进一步的详细说明：
实施例1：
1、称取150g MES于1000mL玻璃瓶中，加入1000mL水溶解，用NaOH调节pH=5.5，将550g EDC和200g NHS加入MES溶液中，完全溶解后，加入磷酸氢二钠60g，磷酸二氢钠40g，调节pH至8.0。
实施例2：
1、称取100g MES于1000mL玻璃瓶中，加入1000mL水溶解，用NaOH调节pH=5.0，将600g EDC和250g NHS加入MES溶液中，完全溶解后，加入磷酸氢二钠30g，磷酸二氢钠20g，调节pH至7.8。

Claims (4)

1.一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法，其特征在于，抗体标记试剂配方包括磷酸氢二钠，磷酸二氢钠，2-吗啉乙磺酸（MES），1-(3-二甲氨基丙基)-3-乙基碳二亚胺（EDC），N-羟基琥珀酰亚胺（NHS）。
2.根据权利要求1所述的一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法，其特征在于磷酸氢二钠3%~6%，磷酸二氢钠2%~5%，2-吗啉乙磺酸（MES）8%~20%，1-(3-二甲氨基丙基)-3-乙基碳二亚胺（EDC）40%~60%，N-羟基琥珀酰亚胺（NHS）15%~30%，总质量组成为100%。
3.根据权利要求1所述的一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法，其特征在于，MES溶液中加入EDC和NHS，用磷酸钠缓冲液调节pH至7.2-9.0。
4.根据权利要求1所述的一种时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法，其特征在于，配制的MES溶液pH=4.6-6.5。
CN201811059144.9A 2018-09-12 2018-09-12 时间分辨荧光免疫层析抗体标记试剂的配方及其制备方法 Pending CN110895282A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111458495A (zh) * 2020-04-07 2020-07-28 上海雄图生物科技有限公司 一种用于时间分辨荧光免疫层析抗体标记试剂及其制备方法

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111458495A (zh) * 2020-04-07 2020-07-28 上海雄图生物科技有限公司 一种用于时间分辨荧光免疫层析抗体标记试剂及其制备方法

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