为了研究我们的位点特异性蛋白质结合策略是否能够有效产生配备有强力细胞毒素的抗体，我们分两步生成了ADC。用LplA W37V催化将曲妥珠单抗（LAP）2用内-BCN b修饰，然后通过SPAAC结合N 3 -PEG 3 -vc - PABC -MMAE偶联高毒性一甲基澳瑞他汀E（MMAE），产生曲妥珠单抗（（ LAP·MMAE）2。接下来，我们评估了曲妥珠单抗（LAP·MMAE）2的体外效能。为此，对SK‐BR‐3细胞和CHO‐K1细胞进行了细胞增殖测定。曲妥珠单抗（LAP·MMAE）2在HER2阳性细胞上显示出强大的亚纳摩尔（IC 50 = 0.31 n m）细胞杀伤特性，而发现该构建体对HER2阴性细胞无害。如预期的那样，曲妥珠单抗（LAP）2缺乏毒素弹头对HER2阳性和阴性细胞均无毒。与目前用于抗体的正交修饰的其他几种抗体标签一样，引入的肽在体内给药后可能具有抗原性。因此，在潜在的医学应用范围内，有必要仔细评估LAP序列的抗原性，这不仅取决于肽序列，还取决于其他因素，例如给药方式或抗体浓度。
通过内-BCN b连接，然后用N 3 -PEG 3 -vc - PABC -MMAE进行SPAAC，生成有效的抗体-药物偶联物曲妥珠单抗（LAP·MMAE）2。上：共轭体系结构。vc =缬氨酸-瓜氨酸，PABC =对氨基苄基氨基甲酸酯，MMAE =单甲基澳瑞他汀E。虚线表示药物释放的可裂解键。剪刀指示溶酶体蛋白酶的切割位点。下图：用曲妥珠单抗（LAP）2和曲妥珠单抗（LAP· MMAE）2（DAR2）处理的SK‐BR‐3和CHO‐K1细胞的细胞增殖测定。图表中的误差线：±1标准偏差。
ADC Linker产品目录：
Fmoc-Val-Cit-PAB CAS:cas:159858-22-7
Fmoc-Val-Cit-PAB-PNP CAS:cas:863971-53-3
Val-cit-PAB-OH CAS:159857-79-1
Phe-Lys(Trt)-PAB CAS:1116085-99-4
Fmoc-Phe -Lys(Trt)-PAB-PNP CAS:1116086-09-9
Aloc-D-Ala-Phe-Lys(Aloc)-PAB-PNP CAS:253863-34-2
Cbz‐Phe‐(Alloc)Lys‐PAB‐PNP CAS:159857-90-6
Boc‐Phe‐(Alloc)Lys‐PAB‐PNP CAS:1160844-44-9
N-SMP CAS:55750‐61‐3
GMBS CAS:80307-12-6
5-Maleimidovalericacid-NHS CAS:103750-03-4
Mal-PEG4-NHS CAS:1325208-25-0
Mal-PEG-NHS CAS:1260092-50-9
5-hexynoic NHS ester CAS:906564-59-8
NHS-PEG4-azide CAS:944251-24-5
Bis-PEG1-NHS ester CAS:65869-64-9
Bis-PEG2-NHS Ester CAS:65869-63-8
Bis-PEG3-NHS Ester CAS:1314378‐16‐9
NHS-PEG3-NHS CAS:1314378-11-4
sulfo-LC-SPDP CAS:150244‐18‐1
MAL-Tetra-EG-OPFP CAS:1347750-84-8
Mal-PEG5-NHS CAS:1315355-92-0
Mal-amido-PEG6-NHS CAS:1137109-21-7
Maleimide-PEG8-NHS ester CAS:756525-93-6
NHS-Ac-N3 CAS:824426-32-6
NHS-PEG12-azide CAS:1610796-02-5
Bis-PEG5-NHS ester CAS:756526-03-1
Bis-PEG9-NHS ester CAS:1008402-79-6
Boc-Val-Cit-PAB-PNP CAS:870487-10-8
Alkyne-PEG-acid (PEG1-PEGn) CAS:55683-37-9
Alkyne-PEG-NHS ester (PEG1-PEGn) CAS:1174157-65-3
Alkyne-PEG-t-butyl ester (PEG1-PEGn) CAS:488150-84-1
Alkyne-PEG-alcohol (PEG1-PEGn) CAS:3973-18-0
Alkyne-PEG-bromide (PEG1-PEGn) CAS:203740-63-0
Alkyne-PEG-Alkyne (PEG1-PEGn) CAS:40842-04-4
Alkyne-PEG2-methylamine CAS:1835759-76-6
Alkyne-PEG4-thiol CAS:1347750-80-4
Alkyne-PEG4-5-nitrophenyl carbonate CAS:1422540-81-5
Alkyne-PEG-methane (PEG1-PEGn) CAS:89635-82-5
Alkyne-PEG4-(CH2)3-methyl ester CAS:1872433-63-0
APN-C3-PEG4-alkyne CAS:2183440-36-8
Alkyne-PEG2-acrylate CAS:52436-42-7
Gly-PEG3-endo-BCN, TFA salt CAS:2354291-37-3
Boc-Gly-PEG3-endo-BCN CAS:2110444-63-6
Aminoethyl-SS-ethylalcohol CAS:15579-01-8
Aminoethyl-SS-propionic acid CAS:15579-00-7
Aminoethyl-SS-ethylamine CAS:51-85-4
t-Boc-Cystamine CAS:485800-26-8
N3-SS-PEG2-acid CAS:2144777-72-8
N3-SS-propionic NHS ester CAS:2243566-44-9
N3-SS-ethylalcohol CAS:1807540-82-4
N3-SS-N3 CAS:352305-38-5
N3-PEG3-SS-PEG3-N3 CAS:1310827-27-0
N3-PEG2-t-Butyl ester CAS:2144777-83-1
Alkyne-PEG1-SS-alcohol CAS:1391914-41-2
Alkyne-PEG1-SS-PEG1-acid CAS:1807503-85-0
Alkyne-PEG1-SS-PEG1-PFP ester CAS:1817735-30-0
Alkyne-PEG1-SS-PEG1-Alkyne CAS:1964503-40-9
Alkyne-PEG1-SS-PEG1-t-butyl ester CAS:1807518-78-0
Acid-PEG-SS-PEG-acid (PEG1-PEGn) CAS:1807539-10-1
Boc-NH-ethyl-SS-propionic acid CAS:485800-27-9
Boc-aminooxy-ethyl-SS-propanol CAS:2128735-21-5
Fmoc-NH-ethyl-SS-propionic acid CAS:864235-83-6
Fmoc-NH-ethyl-SS-propionic NHS ester CAS:2128735-23-7
Mal-NH-ethyl-SS-propionic acid CAS:2128735-24-8
Hydroxy-PEG3-SS-PEG3-alcohol CAS:5662-81-7
THP-SS-alcohol CAS:877864-04-5
THP-SS-PEG1-t-butyl ester CAS:1807503-88-3
THP-SS-PEG1-Tos CAS:1807512-37-3
2-hydroxyethyl disulfide mono-Tosylate CAS:1807530-16-0
Bis-Tos-(2-hydroxyethyl disulfide) CAS:69981-39-1
N3-SS-biotin CAS:1620523-64-9
DBCO-SS-PEG3-biotin CAS:1430408-09-5
Biotin-bisamido-SS-NHS CAS:142439-92-7
NHS-SS-biotin CAS:122266-55-1
Sulfo-NHS-SS-biotin CAS:325143-98-4
PC Biotin-PEG3-alkyne CAS:1869922-24-6
PC Biotin-PEG3-N3 CAS:1937270-46-6
PC Biotin-PEG3-NHS carbonate ester CAS:2353409-93-3
PC Alkyne-PEG4-NHS carbonate ester CAS:1802907-98-7
PC Methyltetrazine-PEG4-NHS carbonate ester CAS:1408057-91-9
PC N3-PEG11-NHS carbonate ester CAS:2353409-91-1
Mal-PEG2-Val-Cit-PAB-OH CAS:2055041-38-6
Mal-PEG1-Val-Cit-PAB-PNP CAS:2249935-92-8
Mal-amido-PEG2-Val-Cit-PAB-PNP CAS:2112738-13-1
N3-PEG3-Val-Cit-PAB-OH CAS:2055024-65-0
N3-PEG3-Val-Cit-PAB-PNP CAS:2055047-18-0
BCN-PEG3-VC-PFP Ester CAS:2353409-45-5
Methylamino-PEG-acid (PEG1-PEGn) CAS:1367918-21-5
小编：axc(西安齐岳生物)