GlpBio

SPDP (SPDP Crosslinker)

目录号: GC30018纯度: >97.50%同义词: 氮-琥珀星氩氨-3(2-吡啶二硫代)-酸酯,SPDP Crosslinker
SPDP (SPDP Crosslinker)是一种短链交联剂，通过NHS酯基团与吡啶二硫反应基团和半胱氨酸硫醇反应，形成可切割（可还原）的二硫键，从而促进胺和硫醇基团的偶联。
SPDP (SPDP Crosslinker)
Cas No.: 68181-17-9
规格价格库存数量操作
1g¥238.00现货
1
5g¥680.00现货
1
10g¥1,250.00现货
1
25g¥2,734.00现货
1
电话: 400-920-5774Email: sales@glpbio.cn

文献被引

本产品暂无引用记录;以下为 GlpBio 产品在 Nature / Cell / Science 等顶刊的客户引用样例
  • Nature cover
    Nature
    641, 529–536 (2025)
  • Nature cover
    Nature
    628, 630–638 (2024)
  • Nature cover
    Nature
    632, 686–694 (2024)
  • Nature cover
    Nature
    618, 1017–1023 (2023)
  • Nature cover
    Nature
    610, 366–372 (2022)
  • Cell cover
    Cell
    187(9):2288-2304 (2024)
  • Cell cover
    Cell
    183(7):1867-1883 (2020)
  • Science cover
    Science
    388(6745) (2025)
  • Science cover
    Science
    387(6739) (2025)
  • Science cover
    Science
    387(6734) (2025)
  • Cell Research cover
    Cell Research
    35, 97–116 (2025)
  • Cell Research cover
    Cell Research
    34, 683–706 (2024)
  • Cell Research cover
    Cell Research
    33, 273–287 (2023)
  • Cell Research cover
    Cell Research
    33, 546–561 (2023)
  • Cell Research cover
    Cell Research
    33, 904–922 (2023)
  • Cell Research cover
    Cell Research
    31, 1291–1307 (2021)

产品描述 Description

SPDP (SPDP Crosslinker) is a short-chain crosslinker that reacts with NHS ester groups and pyridyl disulfide reactive groups with cysteine thiols to form a cleavable (reducible) disulfide bond, thereby facilitating the coupling of amine and thiol groups. When SPDP reacts with free thiols, it releases a detectable byproduct—2-mercaptopyridine—allowing for easy tracking of the reaction by measuring the release of 2-mercaptopyridine at 343 nm. SPDP can be used for the synthesis of antibody-drug conjugates. The SPDP crosslinker is membrane-permeable and can crosslink within cells [1-6].

References:
[1]. Singh V, Mavila AK, et,al. Effect of lysine residue modification of ovine luteinizing hormone by heterobifunctional crosslinking reagent SPDP on subunit-subunit association, receptor binding and biological activity. Indian J Exp Biol. 1992 Nov;30(11):1093-100. PMID: 1284055.
[2]. Lobedanz S, Bokma E et,al. A periplasmic coiled-coil interface underlying TolC recruitment and the assembly of bacterial drug efflux pumps. Proc Natl Acad Sci U S A. 2007 Mar 13;104(11):4612-7. doi: 10.1073/pnas.0610160104. Epub 2007 Mar 5. PMID: 17360572; PMCID: PMC1838649.
[3]. Chen L, Xu N, et,al. Nanoalbumin-prodrug conjugates prepared via a thiolation-and-conjugation method improve cancer chemotherapy and immune checkpoint blockade therapy by promoting CD8+ T-cell infiltration. Bioeng Transl Med. 2022 Jul 30;8(1):e10377. doi: 10.1002/btm2.10377. PMID: 36684090; PMCID: PMC9842047.
[4]. Karumuthil-Melethil S, Perez N, et,al. Dendritic cell-directed CTLA-4 engagement during pancreatic beta cell antigen presentation delays type 1 diabetes. J Immunol. 2010 Jun 15;184(12):6695-708. doi: 10.4049/jimmunol.0903130. Epub 2010 May 14. PMID: 20483724; PMCID: PMC2882504.
[5]. Zhang D, Guo Y, et,al. Expression of a recombinant FLT3 ligand and its emtansine conjugate as a therapeutic candidate against acute myeloid leukemia cells with FLT3 expression. Microb Cell Fact. 2021 Mar 10;20(1):67. doi: 10.1186/s12934-021-01559-6. PMID: 33691697; PMCID: PMC7948335.
[6]. Zhao H, Li L, Peng Z, et,al. Improved targeting delivery of WED-load immunoliposomes modified with SP-A mAb for the treatment of pulmonary fibrosis. Colloids Surf B Biointerfaces. 2023 Apr;224:113237. doi: 10.1016/j.colsurfb.2023.113237. Epub 2023 Mar 1. PMID: 36871414.

SPDP (SPDP Crosslinker)是一种短链交联剂，通过NHS酯基团与吡啶二硫反应基团和半胱氨酸硫醇反应，形成可切割（可还原）的二硫键，从而促进胺和硫醇基团的偶联。当SPDP与游离巯基反应时，能够释放出可检测的副产物—2-巯基吡啶，通过在343nm处测量2-巯基吡啶的释放量可以很容易地跟踪该反应。SPDP可用于合成抗体-药物偶联物。SPDP交联剂具有膜渗透性，可在细胞内交联[1-6]。

实验参考方法 Experimental Reference Method

一、SPDP合成白蛋白偶联物和NanoAlb-proDOX偶联物的方案[1]

1.HSA溶解在PBS-EDTA缓冲液中(20mM sodium phosphate, 150mM NaCl, 1mM EDTA, pH 7.5)，浓度为200μM。

2.在DMSO中制备SPDP原液。在PBS-EDTA缓冲液中，将50μM HSA和1mM SPDP在室温下轻摇2h，以引发反应。

3.反应结束后，通过PD-10柱除去多余的SPDP。

4.DTT溶解在醋酸钠缓冲液中(100mM sodium acetate buffer, 100mM NaCl, pH 4.5) ，终浓度23mg/ml，将DTT溶液加入到第二步反应后的溶液中，使spdp标记的HSA最终浓度约为100μM （醋酸缓冲液与PBS-EDTA缓冲液的比例保持在1:2，以保持反应体系的pH偏酸性，防止DTT还原对天然和细胞内二硫键的影响）。

5. 反应在室温下进行1h，通过PD-10柱除去多余的DTT。

6.将还原后的HSA溶液与100μM的aldoxorubicin （AlDOX，用DMSO总反应体积的20%预稀释）在PBS-EDTA缓冲液中反应，最终HSA浓度为20μM。将混合物在室温下轻轻摇晃一夜。

7.通过PD-10柱流动去除过量的AlDOX。 NanoAlb-proDOX溶液保存在PBS缓冲液中，保存在- 80℃。


二、体内SPDP交联[2]

1.培养物生长到OD600 0.8。

2.样品洗涤两次，用交联缓冲液（20mM Na-phosphate, pH 7.5 /150mM NaCl /1mM EDTA）浓缩7.5倍，在DMSO中分别用DMSO、0.2mM SPDP或0.2mM LC-SPDP孵育30min。

3.用2.5mM Tris·HCl （pH 7.4）淬火后，收获细胞并用交联缓冲液洗涤两次。

4.细胞膜溶解于8M尿素，1% Triton X-100, 20mM Na-phosphate （pH 7.5）中。

5.用50μl Ni-NTA树脂孵育1h，从溶解膜上亲和纯化配合物，首先用10mM imidazole, 8M urea, 1% Triton X-100, 20mM Na-phosphate (pH 7.0)洗涤，然后用20mM imidazole, 1% Triton X-100, 20mM Na-phosphate (pH 7.0), 0.5M NaCl, 0.1% SDS洗涤，最后用30mM imidazole, 1% Triton X-100, 20mM Na-phosphate (pH 7.5), 150mM NaCl, 20% glycerol洗涤。所有的洗涤都进行了三次。

6.用8M urea, 50mM Tris·HCl, 2% SDS, 0.4M imidazole (pH 6.8)洗脱蛋白质，用100mM DTT还原交联剂（37℃，30 min）从络合物中分离， 最后用10% SDS/PAGE分离。

References:
[1]. Chen L, Xu N, et,al. Nanoalbumin-prodrug conjugates prepared via a thiolation-and-conjugation method improve cancer chemotherapy and immune checkpoint blockade therapy by promoting CD8+ T-cell infiltration. Bioeng Transl Med. 2022 Jul 30;8(1):e10377. doi: 10.1002/btm2.10377. PMID: 36684090; PMCID: PMC9842047.
[2]. Lobedanz S, Bokma E, et,al. A periplasmic coiled-coil interface underlying TolC recruitment and the assembly of bacterial drug efflux pumps. Proc Natl Acad Sci U S A. 2007 Mar 13;104(11):4612-7. doi: 10.1073/pnas.0610160104. Epub 2007 Mar 5. PMID: 17360572; PMCID: PMC1838649.

产品文档 Product Documents

Purity:>97.50%

化学性质Chemical Properties

CAS 号
68181-17-9
同义词
氮-琥珀星氩氨-3(2-吡啶二硫代)-酸酯,SPDP Crosslinker
SMILES
O=C(ON1C(CCC1=O)=O)CCSSC2=NC=CC=C2
分子式
C12H12N2O4S2
分子量
312.36 g/mol
溶解性
DMSO : 155 mg/mL (496.22 mM)
保存条件
-20°C, protect from light, stored under nitrogen,unstable in solution, ready to use.
General tips
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。
为了提高溶解度，请将管子加热至 37°C，然后在超声波浴中震荡一段时间。
Shipping Condition
评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备 RT，或根据请求配备蓝冰。

计算工具摩尔浓度 / 稀释 / 分子量 / 单位换算 / 体内配方 / 溶解度

g/mol