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火眼金睛的SEC-MALS:准确测定蛋白分子量,确认蛋白聚体状态

背景
重组蛋白抗原贯穿抗体药物研发的前期免疫、抗体筛选和鉴定、抗体药物候选物体内体外功能验证、抗体药物质量控制以及临床样本分析等过程,而重组蛋白的质量和活性一定程度上直接决定了抗体药物研发的成败和周期。
重组蛋白抗原需要尽可能保持和天然蛋白一致的正确折叠和构象,从而保证蛋白抗原的天然活性,同时具有最小的批间差,帮助提高药物开发成功率。蛋白质的聚集状态是决定其功能和活性的重要性质之一,ACROBiosystems作为药物开发用重组蛋白的行业领导者,致力于“从靶点蛋白开始,为更好生物医药”,不仅在重组蛋白设计、表达和生产过程中严加控制,还建立了准确可靠的分子量测定分析方法来表征重组蛋白在溶液生理条件下的真实聚集状态,指导重组蛋白抗原产品的研发设计、生产工艺优化和质量控制。
蛋白分子量分析技术平台

蛋白分子量分析技术主要有:(1)基于SEC-HPLC(高效分子筛液相色谱)的保留时间内插估算法;(2)基于SEC-MALS(多角度激光散射)的分子量绝对定量法。

两种技术都可以测定溶液中蛋白的分子量,但在原理上有一些不同,两种技术的异同点比较如下:

检测方法 原理和特点 结果
SEC-HPLC 柱校正法 通过比较待测蛋白和标准蛋白的洗脱保留时间计算相对分子量
假设待测蛋白与标准蛋白的构象,形状相似
假设与柱填料无任何非特异的相互作用
假设每次洗脱时间可重现无漂移
SEC-MALS 光散射法 通过激光散射强度直接测定绝对分子量,分子量测定不依赖于洗脱体积或保留时间
散射光强度正比于摩尔质量、浓度、折光指数增量的平方
散射光角度依赖性仅与分子尺寸大小相关
SEC-MALS技术基本原理
MALS技术是基于蛋白的分子量与光散射的强度直接相关来测定蛋白绝对分子量的技术,无需使用标准蛋白,不依赖洗脱体积。通过SEC分离后的各组分进入多角光散射检测器,激光照射到分析物时会发生光的散射,MALS通过多个角度同时测定散射光的强度(Fig1.a),公式b1可以看出散射光强度正比于摩尔质量、浓度、折光指数增量的平方,且通过用公式b2可以直接计算出分析物的绝对分子量。
因此,在生物制药领域,SEC-MALS广泛应用于研发和分析实验,表征蛋白质和聚合物、评估溶液特性、工艺开发和质量控制等。

Fig 1. SEC-MALS检测原理图 a.光散射示意图 b.分析物分子量计算

通过HPSEC-MALS测定HIV-1包膜三聚体的分子量,结果可靠性经正交质谱验证
美国国家过敏和传染病研究所疫苗研究中心疫苗生产计划实验室研究人员[1]利用HPSEC-MALS快速稳定地测定了高糖基化HIV-1包膜三聚体——Trimer 4571 疫苗的总分子量(339KDa)以及蛋白质(213KDa)和聚糖组分(126KDa)的分子量,测定结果也进一步经正交质谱分析验证,这有助于支持工艺开发并提供快速的蛋白质表征数据,也证明HPSEC-MALS对许多正在开发的异源糖蛋白候选疫苗具有广泛的适用性。

Fig 2. SEC-MALS 获得的 Trimer 4571 分子量结果与 MS数据的比较

Fig 3. 三聚体的SEC-UV-dRI-MALS测定结果

以上市药物抗体分子量测定为例,比较SEC-HPLC和SEC-MALS的测定结果
我们用SEC-HPLC和MALS两种方法分别测定了OKT3、赫赛汀(Herceptin)和伊匹单抗(Ipilimumab)三种已知分子量的上市抗体药物。如Tab 1所示,三种抗体药物实际具有非常接近的分子量(150-160kDa),但三者洗脱保留时间却有较大的差异,造成SEC-HPLC计算分子量大相径庭,其中OKT3抗体测定为212.5kDa,Ipilimumab仅为66.3kDa,和真实分子量相去甚远。而SEC-MALS检测三种抗体分子量和抗体理论分子量非常接近(Fig 5)。
由此可见,MALS技术测定蛋白分子量的结果较传统的SEC-HPLC更加准确可靠。
Tab 1. 三种抗体药分子量测定结果
抗体名称 Column 保留时间
TR (min)
分子量
(SEC-HPLC)
分子量
(SEC -MALS)
OKT3  G3000SWxl 14.775 212.5 kDa  161.6 kDa
Herceptin G3000SWxl  15.436 146 kDa  151 kDa 
lpilimumab G3000SWxl  17.039 66.3 kDa  172.2 kDa

Fig 4. 不同抗体的SEC-HPLC分子量测定结果

Fig 5. SEC-MALS测定三种抗体药物的分子量

火眼金睛的SEC-MALS:确保多聚体蛋白产品的质量
天然蛋白在体内往往以一定的聚集状态和构象存在,保证其发挥正确的生物学功能,而重组蛋白在生产过程中产生的随机不正确聚集常常会引起某些关键功能表位构象或暴露程度的改变,导致无法获得针对天然有效表位的高质量抗体候选物,影响抗体药物体外功能验证等结果,显著降低抗体药物开发的成功率。
ACROBiosystems深谙重组蛋白结构对其功能的影响,充分了解抗体药物开发的需求,深入研究复合物的结构性质,采用特有的技术平台设计开发了多种同源/异源多聚体产品,并用SEC-MALS对产品分子量进行验证,将SEC-MALS纯度结果作为质控标准,确保产品性质更均一,批间差更小,同时,产品活性经过ELISA、流式细胞,功能等应用场景的验证,加速抗体药物的开发。

案例一:TNFSF三聚体蛋白

肿瘤坏死因子(TNF)超家族系列蛋白在天然状态下为三聚体结构,只有当三聚体形式TNF-α与TNFR1的胞外区结合时才能引起TNFR1的三聚化,进而有效的激发下游通路引起细胞凋亡。
SEC-MALS验证蛋白结构和纯度
Cat.No. OXL-H52Q8(Human OX40 Ligand /TNFSF4 Protein, His Tag, active trimer)

Fig 6. 经MALS验证,人源 OX40 Ligand, His Tag, active trimer (Cat. No. OXL-H52Q8)的纯度超过 95%,分子量约为60-80kDa。

FACS验证蛋白细胞功能活性
Cat.No. OXL-H52Q8(Human OX40 Ligand /TNFSF4 Protein, His Tag, active trimer)

Fig 7. FACS 分析显示,人源 OX40 Ligand, His Tag (active trimer) (Cat. No. OXL-H52Q8)可以与过表达人源OX40的 293T细胞结合。所用 OX40 Ligand浓度为0.1 μg/mL。

Protocol

案例二:IL-2R 异源二聚/三聚体

IL-2的受体(IL-2R)包含α、β和γ三个亚单位。三个亚单位可以组成不同亲和力的受体。由于IL-2受体复合物结构的特殊性,在没有IL-2存在时,α、β和γ无法直接形成高亲和力的IL-2受体复合物。ACROBiosystems利用自有专利技术开发了一系列IL-2 R alpha & IL-2 R beta & IL-2 R gamma异源三聚体蛋白和IL-2 R beta & IL-2 R gamma异源二聚体蛋白。
SEC-MALS验证蛋白结构和纯度
Cat.No. ILG-H5254(Human IL-2RB&IL-2RG Heterodimer Protein, Fc Tag&Fc Tag)

Fig 8. 经MALS验证,人源 IL-2RB&IL-2RG 异源二聚体蛋白, Fc Tag&Fc Tag(Cat. No.ILG-H5254)的纯度超过90%,且蛋白的分子量在145-165 kDa左右。

Cat. No. ILG-H5257(Human IL-2RB&IL-2RA&IL-2RG, Fc Tag&Fc Tag)

Fig 9. 经MALS验证,人源 IL-2RB&IL-2RA&IL-2RG异源三聚体蛋白, Fc Tag&Fc Tag(Cat. No. ILG-H5257)的纯度超过90%,且该蛋白的分子量在175-190 kDa左右。

经SPR验证,IL-2与IL-2受体的亲和力验证结果与文献[2]基本一致

Fig 10. IL-2 与 ACRO 的 IL2R 构建体的结合亲和力经SPR验证。

案例三:CTLA-4同源共价二聚体

随着双特异性药物的兴起,基于PD-1/PD-L1以及CTLA-4双靶点又引起了一波双抗药物研发热潮。目前已经有多种类似的双抗药物进入临床验证阶段,其适应症也覆盖了多种实体瘤。CTLA-4(细胞毒性T淋巴细胞相关蛋白4),也称为CD152(分化簇 152),是T细胞的一种蛋白受体,通常以同源二聚体形式存在。
SEC-MALS验证蛋白结构和纯度
Cat. No. CT4-H52H9(Human CTLA-4 / CD152 Protein, His Tag, active dimer (MALS verified))

Fig 11. 经 SEC-MALS 验证,人源CTLA-4, His Tag (Cat. No. CT4-H52H9) 的纯度超过90%,且该蛋白的分子量约为 45-60 kDa。

SPR验证CTLA-4二聚体亲和力

Fig 12. 经SPR验证,CTLA-4二聚体亲和力比单体提高10倍以上

案例四:Spike蛋白三聚体

S蛋白是新冠病毒研究中最重要的靶标,在病毒表面以同源三聚体形式存在,当S1亚基中的RBD与宿主细胞ACE2结合后,引发三聚体不稳定性,造成S1亚基和S2亚基发生解离,S2亚基的构象变为高度稳定的融合后结构,因此S蛋白具有融合前(Prefusion)和融合后(Postfusion)两种构象。研究人员发现,S蛋白的构象形式很不稳定,即使不与宿主细胞受体ACE2结合,也会自发出现由Prefusion到Postfusion的构象改变,这对疫苗开发和中和性抗体筛选带来巨大的挑战。
SEC-MALS&NS-EM验证蛋白结构和纯度

Cat. No. SPN-C52H9(SARS-CoV-2 S protein, His Tag, Super stable trimer (MALS & NS-EM verified))

Fig 13. 经还原条件SDS-PAGE验证,SARS-CoV-2 S 蛋白、His Tag、Super stable trimer (MALS & NS-EM verified) (Cat. No. SPN-C52H9)的纯度通过超过90%,经SEC-MALS验证分子量约为550-660 kDa。大小和外观与NS-EM验证与已发表文献中报道的 SARS-CoV-2 三聚体相似。

S蛋白三聚体可用于检测康复病人血清样品抗体滴度,信噪比高

Fig 14. S-6P蛋白,S-2P蛋白与S蛋白检测康复病人血清样本的抗体滴度的对比分析。结果发现,S-6P的信噪比显著高于S蛋白,S-2P以及其它公司的S蛋白,说明S-6P更适合于进行血清样本的抗体滴度检测,能在注射疫苗后的早期从血清中检测出抗体滴度水平,有利于疫苗的免疫原性评价。

产品列表
  • TNFSF
  • CD3系列
  • FcRn (FCGRT & B2M)
  • Interleukin
  • Spike trimer
  • 其他
分子 货号 产品描述
4-1BB Ligand41L-C5254Cynomolgus 4-1BB Ligand / TNFSF9 Protein, Fc Tag, active (MALS verified)
41L-H5265Human 4-1BB Ligand / TNFSF9 (71-254) Protein, Fc Tag, active (MALS verified)
41L-H52D4Human 4-1BB Ligand / TNFSF9 (71-254) Protein, His,Flag Tag (active ) (MALS verified)
APRILAPL-C52D2Cynomolgus APRIL / TNFSF13 Protein, His,Flag Tag, active (MALS verified)
APL-H5268Human APRIL / TNFSF13 Protein, Fc Tag, active (MALS verified)
APL-H52D1Human APRIL / TNFSF13 Protein, His,Flag Tag, active (MALS verified)
BAFFBAF-H5261Human BAFF / TNFSF13B / CD257 Protein, Fc Tag, active (MALS verified)
BAF-H52D4Human BAFF / TNFSF13B / CD257 Protein, His,Flag Tag, active (MALS verified)
BAF-H82F3Biotinylated Human BAFF / TNFSF13B / CD257 Protein, Avitag™,Fc Tag (active ) (MALS verified)
BAF-H82Q2Biotinylated Human BAFF / TNFSF13B / CD257 Protein, His,Avitag™ (active ) (MALS verified)
CD27 LigandCDD-HF2D4FITC-Labeled Human CD27 Ligand / CD70 Protein, His,Flag Tag active (MALS verified)
CDL-H5266Human CD27 Ligand / CD70 Protein, Fc Tag (active ) (MALS verified)
CDL-H52DaHuman CD27 Ligand / CD70 Protein, His,Flag Tag (active ) (MALS verified)
CDL-H82D7Biotinylated Human CD27 Ligand / CD70 Protein, His,Avitag™,Flag Tag (active ) (MALS verified)
CDL-M5245Mouse CD27 Ligand / CD70 Protein, His Tag (active ) (MALS verified)
CD40 LigandCDL-H5269Human CD40 Ligand / TNFSF5 Protein, Fc Tag (active ) (MALS verified)
CDL-H52DbHuman CD40 Ligand / TNFSF5 Protein, His,Flag Tag (active ) (MALS verified)
CDL-H82DbBiotinylated Human CD40 Ligand / TNFSF5 Protein, Avitag™,His,Flag Tag (active ) (MALS verified)
CDL-M5248Mouse CD40 Ligand / TNFSF5 Protein, His Tag (active ) (MALS verified)
CDL-M52D5Mouse CD40 Ligand / TNFRSF5 Protein, His,Flag Tag (active ) (MALS verified)
CDL-M82H5Biotinylated Mouse CD40 Ligand / TNFSF5 Protein, His,Avitag™ (active ) (MALS verified)
Fas LigandFAL-H5241Human Fas Ligand / TNFSF6 Protein, His Tag, active (MALS verified)
LIGHTLIT-H5242Human LIGHT / TNFSF14 Protein, His Tag (active ) (MALS verified)
LIT-H5269Human LIGHT / TNFSF14 Protein, Fc Tag (active ) (MALS verified)
OX40 LigandOXL-H5266Human OX40 Ligand / TNFSF4 Protein, Fc Tag (active ) (MALS verified)
OXL-H52Q8Human OX40 Ligand / TNFSF4 Protein, His Tag (active ) (MALS verified)
TNF-alphaTNA-C52H3Canine TNF-alpha Protein, His Tag, active (MALS verified)
TNA-H4211ActiveMax® Human TNF-alpha Protein, Tag Free, low endotoxin (active ) (MALS verified)
TNA-H5228Human TNF-alpha Protein, His Tag (active ) (MALS verified)
TNA-H8211Biotinylated Human TNF-alpha Protein, epitope tag free, ultra sensitivity (primary amine labeling) (active ) (MALS verified)
TNA-H82E3Biotinylated Human TNF-alpha Protein, His,Avitag™ (active ) (MALS verified)
TNA-M82E9Biotinylated Mouse TNF-alpha Protein, His,Avitag™ (active ) (MALS verified)
TNFSF11RAL-H5240Human TNFSF11 / RANKL / CD254 Protein, His Tag (active ) (MALS verified)
RAL-H82Q5Biotinylated Human TNFSF11 / RANKL / CD254 Protein, His,Avitag™, active (MALS verified)
分子 货号 产品描述
CD3E & CD3DCDD-C52W4Cynomolgus CD3 epsilon&CD3 delta Protein, His Tag&Flag Tag (MALS verified)
CDD-C52W9Cynomolgus CD3 epsilon&CD3 delta Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDD-C82W6Biotinylated Cynomolgus CD3E&CD3D Protein, His,Avitag™&Tag Free (MALS verified)
CDD-H5255Human CD3 epsilon&CD3 delta Protein, Fc Tag&Fc Tag (MALS verified)
CDD-H52W1Human CD3 epsilon & CD3 delta Protein, His Tag&Tag Free (MALS verified)
CDD-H52WaHuman CD3 epsilon&CD3 delta Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDD-H82F6Biotinylated Human CD3 epsilon&CD3 delta Protein, Fc,His,Avitag™&Fc,Flag,Avitag™ (MALS verified)
CDD-H82W0Biotinylated Human CD3E&CD3D Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling) (MALS verified)
CDD-H82W6Biotinylated Human CD3 epsilon&CD3 delta Protein, His,Avitag™&Tag Free (MALS verified)
CDD-M5219Mouse CD3 epsilon&CD3 delta Protein, His Tag&Flag Tag (MALS verified)
CDD-M52W2Mouse CD3 epsilon&CD3 delta Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDD-R5257Rat CD3 epsilon&CD3 delta Protein, Llama Fc Tag&Llama Fc Tag (MALS verified)
CDD-R5259Rabbit CD3E&CD3D Protein, Llama Fc&Llama Fc, low endotoxin (MALS verified)
CDD-R52D7Rabbit CD3 epsilon&CD3 delta Protein, His Tag&Flag Tag (MALS verified)
CDD-R52D8Rat CD3 epsilon&CD3 delta Protein, His Tag&Flag Tag (MALS verified)
CD3E & CD3GCDG-C5257Cynomolgus CD3E&CD3G Protein, Fc Tag&Fc Tag (MALS verified)
CDG-C52W6Cynomolgus CD3E&CD3G Protein, His Tag&Tag Free (MALS verified)
CDG-C8219Biotinylated Cynomolgus CD3E&CD3G Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-H5253Human CD3 epsilon&CD3 gamma Protein, Fc Tag&Fc Tag (MALS verified)
CDG-H52W5Human CD3 epsilon&CD3 gamma Protein, His Tag&Tag Free (MALS verified)
CDG-H52W9Human CD3 epsilon&CD3 gamma Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CDG-H82W3Biotinylated Human CD3 epsilon&CD3 gamma Protein, His,Avitag™&Tag Free (MALS verified)
CDG-M52D2Mouse CD3 epsilon&CD3 gamma Protein, His Tag&Flag Tag (MALS verified)
CDG-M52W2Mouse CD3 epsilon&CD3 gamma Protein,Fc,His Tag&Fc,Flag Tag (MALS verified)
CDG-M58D2Biotinylated Mouse CD3 epsilon&CD3 gamma Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-R82D3Biotinylated Rabbit CD3 epsilon&CD3 gamma Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-R82D4Biotinylated Rat CD3 epsilon&CD3 gamma Protein, His,Avitag™&Flag Tag (MALS verified)
CDG-RB52D5Rabbit CD3 epsilon&CD3 gamma Protein, His Tag&Flag Tag (MALS verified)
分子 货号 产品描述
FcRn (FCGRT & B2M)FCM-C5284Cynomolgus / Rhesus macaque FcRn / FCGRT&B2M Protein, His Tag&Strep II Tag (MALS & BLI verified)
FCM-C52W9Cynomolgus / Rhesus macaque FcRn / FCGRT&B2M Protein, His Tag (MALS & BLI verified)
FCM-C82W3Biotinylated Cynomolgus / Rhesus macaque FcRn / FCGRT&B2M Protein, His,Avitag™ (MALS & SPR verified)
FCM-H5286Human FcRn / FCGRT&B2M Protein, His Tag&Strep II Tag (SPR & BLI & MALS verified)
FCM-H82W4Biotinylated Human FcRn / FCGRT&B2M  Protein, Avitag™,His Tag&Strep II Tag (SPR & BLI & MALS verified)
FCM-H82W7Biotinylated Human FcRn / FCGRT&B2M Protein, His,Avitag™ (MALS & SPR verified)
FCM-M52W2Mouse FcRn / FCGRT&B2M Protein, His Tag&Twin-Strep Tag (MALS & BLI verified)
FCM-M52W8Mouse FcRn / FCGRT&B2M Protein, His Tag (MALS & BLI verified)
FCM-M82W5Biotinylated Mouse FcRn / FCGRT&B2M Protein, His,Avitag™ (MALS & BLI verified)
FCM-M82W6Biotinylated Mouse FcRn / FCGRT&B2M Protein, Avitag™,His Tag&Twin-Strep Tag (MALS & BLI-verified)
FCM-P5280Porcine FcRn / FCGRT&B2M Protein, His Tag&Strep II Tag (MALS & SPR verified)
FCM-R5287Rat FcRn / FCGRT&B2M Protein, His Tag&Strep II Tag (MALS & SPR verified)
FCM-R52W5Rabbit FcRn / FCGRT&B2M Protein, His Tag&Tag Free (MALS & SPR verified)
FCM-R82W7Biotinylated Rat FcRn / FCGRT&B2M Protein, Avitag™,His Tag&Strep II Tag (MALS & SPR verified)
FCN-B82W3Biotinylated Bovine FcRn / FCGRT&B2M Protein, His,Avitag™ (MALS & SPR verified)
FCN-F82W3Biotinylated Feline FcRn / FCGRT&B2M Protein, His,Avitag™ (MALS & SPR verified)
FCN-H52W7Human FcRn / FCGRT&B2M Protein, His Tag (SPR & BLI & MALS verified)
分子 货号 产品描述
IL-17A & IL-17FILF-H52W6Human IL-17A&IL-17F Protein, Twin Strep&His Tag (MALS verified)
IL-2 R beta & IL-2 R gammaILG-C82F4Biotinylated Cynomolgus IL-2 R beta&IL-2 R gamma Protein, Fc,Avitag™&Fc,Avitag™ (MALS verified)
ILG-H5254Human IL-2 R beta&IL-2 R gamma Protein, Fc Tag&Fc Tag (MALS verified)
ILG-H5283Human IL-2 R beta&IL-2 R gamma Protein, His Tag&Twin-Strep Tag (MALS verified)
ILG-H82F3Biotinylated Human IL-2 R beta&IL-2 R gamma Protein, Fc,Avitag™&Fc,Avitag™ (MALS verified)
ILG-M5254Mouse IL-2 R beta&IL-2 R gamma Protein, Fc Tag&Fc Tag (MALS verified)
ILG-M5283Mouse IL-2 R beta&IL-2 R gamma Protein, His Tag&Twin-Strep Tag (MALS verified)
ILG-R5283Rat IL-2 R beta&IL-2 R gamma Protein, His Tag&Twin-Strep Tag (MALS verified)
IL-7 R alpha & TSLP RILR-H5255Human IL-7 R alpha&TSLP R Protein, Fc Tag&Fc Tag (MALS verified)
IL23A & IL12BILB-HR52W3Human IL-23 alpha & Rat IL-12 beta Protein, His Tag&Tag Free (MALS verified)
分子 货号 产品描述
Spike ProteinSPN-C52H3SARS-CoV-2 S protein (D614G), His Tag, Super stable (MALS verified)
SPN-C52H7SARS-CoV-2 S protein, His Tag, Super stable (MALS verified)
SPN-C52H9SARS-CoV-2 S protein, His Tag, Super stable (MALS & NS-EM verified)
SPN-C52HaSARS-CoV-2 Spike (T95I, Y144S, Y145N, R346K, E484K, N501Y, D614G, P681H, D950N), His Tag (MALS verified)
SPN-C52HbSARS-CoV-2 Spike (C136F, Y144del, R190S, D215G, LA242-243del, Y449H, E484K, N501Y, D614G, H655Y, N679K, T716I, T859N), His Tag (MALS verified)
SPN-C52HeSARS-CoV-2 Spike (T19R, G142D, EF156-157del, R158G, L452R, T478K, D614G, P681R, D950N), His Tag (MALS verified)
SPN-C52HhSARS-CoV-2 Spike (P26S, HV69-70del, V126A, Y144del, LLA241-243del, H245Y, S477N, E484K, D614G, P681H, T1027I, D1118H), His Tag (MALS verified)
SPN-C52HkSARS-CoV-2 S protein (L18F, D80A, D215G, 242-244del, R246I, K417N, E484K, N501Y, D614G, A701V) , His Tag (MALS verified)
SPN-C52HmSARS-CoV-2 Spike (K417N, D614G), His Tag (MALS verified)
SPN-C52HnSARS-CoV-2 Spike (K417T, D614G), His Tag (MALS verified)
SPN-C52HpSARS-CoV-2 Spike (N501Y, D614G), His Tag (MALS verified)
SPN-C52HqSARS-CoV-2 Spike (E484K, D614G), His Tag (MALS verified)
SPN-C52HrSARS-CoV-2 Spike (T95I, G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H), His Tag (MALS verified)
SPN-C52HsSARS-CoV-2 Spike (G75V, T76I, SYLTPGD 247-253 del, L452Q, F490S, D614G, T859N), His Tag (MALS verified)
SPN-C52HtSARS-CoV-2 Spike (T19R, V70F, FR157-158Del, A222V, W258L, K417N, L452R, T478K, D614G, P681R, D950N), His Tag (MALS verified)
SPN-C52HvSARS-CoV-2 Spike (D614G), His Tag (MALS verified)
SPN-C52HzSARS-CoV-2 Spike , His Tag (B.1.1.529/Omicron) (MALS verified)
SPN-C82E3Biotinylated SARS-CoV-2 S protein (D614G), His,Avitag™, Super stable (MALS verified)
SPN-C82E4Biotinylated SARS-CoV-2 S protein (L18F, D80A, D215G, 242-244del, R246I, K417N, E484K, N501Y, D614G, A701V) , His,Avitag™ (MALS verified)
SPN-C82E5Biotinylated SARS-CoV-2 S protein (HV69-70del, Y144del, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H) , His,Avitag™ (MALS verified)
SPN-C82E6Biotinylated SARS-CoV-2 S protein (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, V1176F) , His,Avitag™ (MALS verified)
SPN-C82E7Biotinylated SARS-CoV-2 Spike (T95I, G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H), His,Avitag™ (MALS verified)
SPN-C82E9Biotinylated SARS-CoV-2 S protein, His,Avitag™, Super stable (MALS verified)
SPN-C82EcBiotinylated SARS-CoV-2 Spike (T19R, G142D, EF156-157del, R158G, L452R, T478K, D614G, P681R, D950N), His,Avitag™ (MALS verified)
SPN-C82EdBiotinylated SARS-CoV-2 Spike (P26S, HV69-70del, V126A, Y144del, LLA241-243del, H245Y, S477N, E484K, D614G, P681H, T1027I, D1118H), His,Avitag™ (MALS verified)
SPN-C82EeBiotinylated SARS-CoV-2 Spike , His,Avitag™ (B.1.1.529/Omicron) (MALS verified)
SPN-C82EhBiotinylated SARS-CoV-2 Spike (G75V, T76I, SYLTPGD 247-253 del, L452Q, F490S, D614G, T859N), His,Avitag™ (MALS verified)
SPN-C82EiBiotinylated SARS-CoV-2 Spike (T19R, V70F, FR157-158Del, A222V, W258L, K417N, L452R, T478K, D614G, P681R, D950N), His,Avitag™ (MALS verified)
SPN-C82EkBiotinylated SARS-CoV-2 Spike (T95I, Y144S, Y145N, R346K, E484K, N501Y, D614G, P681H, D950N), His,Avitag™ (MALS verified)
SPN-C82EqBiotinylated SARS-CoV-2 Spike (C136F, Y144del, R190S, D215G, LA242-243del, Y449H, E484K, N501Y, D614G, H655Y, N679K, T716I, T859N), His,Avitag™ (MALS verified)
SPN-H52H5HCoV-HKU1 (isolate N5) Spike , His Tag (MALS verified)
SPN-H82E3Biotinylated HCoV-HKU1 (isolate N5) Spike , His,Avitag™ (MALS verified)
SPN-M52H4MERS Spike protein (R748A, R751A, V1060P, L1061P), His Tag (MALS verified)
Spike RBDSPD-C52M5SARS-CoV-2 S protein RBD , His Tag (MALS verified)
分子 货号 产品描述
CD28CD8-H525aHuman / Cynomolgus / Rhesus macaque CD28 Protein, Fc Tag, active (MALS verified)
CD8-H52HcHuman / Cynomolgus / Rhesus macaque CD28 Protein, His Tag, active (MALS verified)
CD8-H82E5Biotinylated Human / Cynomolgus / Rhesus macaque CD28 Protein, His,Avitag™, active (MALS verified)
CD8-H82F2Biotinylated Human / Cynomolgus / Rhesus macaque CD28 Protein, Fc,Avitag™, active (MALS verified)
CD8-M52H6Mouse CD28 Protein, His Tag, active (MALS verified)
CD8-M82E3Biotinylated Mouse CD28 Protein, His,Avitag™, active (MALS verified)
CD79A & CD79BCDB-H52W3Human CD79A&CD79B Hetero Protein, Mouse IgG2a Fc,Flag Tag&Mouse IgG2a Fc,His Tag (MALS verified)
CD8 alpha & betaCDA-H52W5Human CD8 alpha&beta (CD8A&CD8B) Hetero Protein, His Tag&Tag Free (MALS verified)
CTLA-4CT4-H52H9Human CTLA-4 / CD152 Protein, His Tag, active (MALS verified)
DPPIVDP4-H5211Human DPPIV / CD26 Protein, Tag Free, active (active enzyme, MALS verified)
FAPFAP-C52H3Cynomolgus FAP Protein, His Tag (MALS verified)
FAP-C82Q5Biotinylated Cynomolgus FAP Protein, His,Avitag™ (MALS verified)
FAP-H5244Human FAP Protein, His Tag (MALS verified) (active enzyme)
FAP-H5263Human FAP Protein, Fc Tag (active enzyme, MALS verified)
FAP-H82Q6Biotinylated Human FAP Protein, His,Avitag™ (MALS verified)
FAP-HF243FITC-Labeled Human FAP Protein, His Tag (MALS verified)
FAP-M52H3Mouse FAP Protein, His Tag (MALS verified)
FAP-M82Q5Biotinylated Mouse FAP Protein, His,Avitag™ (MALS verified)
Integrin alpha 2 beta 1IT1-M52W3Mouse Integrin alpha 2 beta 1 (ITGA2&ITGB1) Hetero Protein, His Tag&Tag Free (MALS verified)
LRRC32 & TGF-beta 1GA1-H52W9Human LRRC32&TGF-beta 1 Hetero protein, His Tag&Tag Free (MALS verified)
GA1-M52W2Mouse LRRC32&TGF-beta 1 Hetero protein, His Tag&Tag Free (MALS verified)
PSMAPSA-C5247Cynomolgus PSMA / FOLH1 Protein, His Tag (MALS verified) (active enzyme)
PSA-H52H3Human PSMA / FOLH1 Protein, His Tag (active enzyme, MALS verified)
PSA-H82QbBiotinylated Human PSMA / FOLH1 Protein, His,Avitag™ (active enzyme, MALS verified)
PSA-R5245Rat PSMA / FOLH1 Protein, His Tag (active enzyme, MALS verified)
TIGITTIT-C5253Cynomolgus / Rhesus macaque TIGIT Protein, Mouse IgG2a Fc Tag (MALS verified)
TIT-C52H7Canine TIGIT Protein, His Tag (MALS verified)
TIT-H5254Human TIGIT Protein, Fc Tag (MALS verified)
TIT-H52H5Human TIGIT Protein, His Tag (MALS verified)
TIT-H82E6Biotinylated Human TIGIT Protein, His,Avitag™ (recommended for biopanning) (MALS verified)
TIT-H82F1Biotinylated Human TIGIT Protein, Fc,Avitag™ (MALS verified)
TIT-H82F3Biotinylated Human TIGIT Protein, Mouse IgG2a Fc,Avitag™ (MALS verified)
TIT-M5253Mouse TIGIT Protein, Mouse IgG2a Fc Tag (MALS verified)
TIT-M52E6Mouse TIGIT Protein, His Tag (MALS verified)

参考文献

  • [1]Bender M F , Li Y , Ivleva V B , et al. Protein and glycan molecular weight determination of highly glycosylated HIV-1 envelope trimers by HPSEC-MALS[J]. Vaccine, 2021(7523).
  • [2]Sriram B , Tania C R , Davis A M , et al. Ligand binding kinetics of IL-2 and IL-15 to heteromers formed by extracellular domains of the three IL-2 receptor subunits[J]. International Immunology(11):1839.

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