Claudin 18.2 Proteins & Stable Cell Lines

Products

Recombinant Proteins

• Active membrane protein
• Consistent Supply

Stable Cell Lines

• Single-clone derived
• Overexpression guarantee
• Stable over 15 passages

Applications

Product Performance

• Product Application Validations
  

  Figure 2.  Bioactivity of CLDN18.2, hFc, Human (Cat. No.: Z03709) was validated by ELISA.

  Immobilized anti-Claudin 18.2 antibody was at 1 μg/mL (100 μL/well). Dose response curve for CLDN18.2, hFc, Human with the EC50 of 17.60 ng/ml was determined by ELISA. CLDN18.2, hFc, Human was found to be an active membrane protein.  

  

  Figure 3. Cell-based binding assay with CHO-K1/CLDN 18.2 stable cell line (Cat. No.: M00916).

  CHO-K1/CLDN18.2 cells were plated at 5x10⁵ cells/well in 100 µL of PBS in 96-well plate, incubated with gradient concentration of IMAB-362 (anti-CLDN18.2 antibody) on ice for 1 hour. The supernatant was discarded and the cells were incubated with goat anti-human IgG antibody at 10 µg/mL on ice for 1 hour. The mean fluorescence intensity of APC (Mean APC-H) was analyzed in live cell gate by a flow cytometer .

  

  Figure 4. Cell based ELISA with CHO-K1/CLDN 18.2 cell line (Cat. No.: M00916).

  CHO-K1/CLDN18.2 cells were plated at 8x10⁴ cells /well in 100 μl DMEM plus with 10% FBS in 96-well plate overnight. Then the cells were fixed with paraformaldehyde for 15 minutes. After washing, the cells were incubated with gradient concentration of IMAB-362 (anti-CLDN18.2 antibody) for 1 hour, and incubated with goat anti-human IgG (H&L) [HRP] antibody. After incubation with TMB substrate, OD450 was measured by a microplate reader.

• Product Characterizations
  

  Figure 5.  The purity of CLDN18.2, His, Human (Cat. No.: Z03504) was assessed by SDS-PAGE. Purity > 90% was observed for 2 μg of protein.

  Lane BSA: 2 μg BSA.
  Lane R: 2 μg CLDN18.2, reducing (R), no heat treatment before loading.

  

  Figure 6. Protein expression of CLDN18.2 was validated in HEK293/CLDN18.2 stable cell line (Cat. No.: M00917) using FACS. The FACS analysis of CLDN18.2 expression in HEK293/ CLDN18.2 clone was conducted using anti-CLDN18.2 antibody. The cells analyzed were live cells (E1 gate).

  Green line: negative control, treated with goat anti-human IgG antibody.
  Red line: tested sample, treated with anti-CLDN18.2 antibody.

  

  Figure 7. Stability of CLDN18.2 expression in HEK293/CLDN18.2 stable cell line (Cat. No.: M00917) was tested using FACS. The stability of expression was tested for cell samples from 20 passages using anti-CLDN18.2 antibody. The cells analyzed were live cells (E1 gate).

  Blue line: blank control, untreated cells.
  Purple line: negative control, treated with goat anti-human IgG antibody.
  Experimental group: cell samples of different passages.

Related Products

CLDN18.2 Clinical Trials and Therapies

• Claudin 18.2 - Targeted Drugs and Therapies

  At present, four main types of Claudin 18.2 targeted therapies have been approved for clinical trials - monoclonal antibodies, CAR-T cell therapy, bispecific antibodies and ADCs. Among these, anti-CLDN18.2 monoclonal antibody therapy represents the most widely adopted therapeutic approach using Claudin 18.2. One of the challenges in the development of a targeted monoclonal antibody is to design an antibody that differentiates between the isoforms by specifically recognizing Claudin 18.2 but not Claudin 18.1. This is difficult to do because the only difference between these two splice variants is a change of 7-8 amino acids in the first extracellular loop ^[12]. Having a high-quality Claudin 18.2 antigen can help pharmaceutical companies achieve breakthrough using this target.

  IMAB362 (also known as Zolbetuximab and Claudiximab) developed by Ganymed Pharmaceuticals AG, is a Claudin 18.2 antibody drug that has entered Phase III of clinical trial. It is a novel chimeric IgG1 antibody highly specific to Claudin 18.2. It binds to Claudin 18.2 on the tumor cell surface and stimulates cellular and soluble immune effectors that activate antibody-dependent cytotoxicity (ADCC) and complement dependent cytotoxicity (CDC) ^[11].

  CAR-T cells can also recognize Claudin 18.2 expressed on tumor cells, and directly attack these tumor cells. In May of 2020, the world's first CAR-T therapy targeting Claudin 18.2 CT-041 got FDA’s IND clearance. Results from phase I clinical trial published in 2019 show a total objective response rate of 33% ^[13].

  In June of 2020, the world’s first Claudin18.2/PD-L1 bispecific antibody preclinical data was published on AACR. In July of 2020, Amgen received the first approval of CD3/Claudin 18.2 bispecific antibody AMG 910 to enter clinical stage.

• Table 1. Claudin 18.2 drugs under development

  Drug Name	Target	Types	Phase	Indications	Company

  zolbetuximab	Claudin 18.2; Complement cascade	Monoclonal Antibody	Clinical Phase III	Metastatic esophageal cancer; Metastatic pancreatic cancer; Metastatic gastric cancer	Astellas Pharma
  CT-041	Claudin 18.2	CAR-T	Clinical Phase II	Gastric adenocarcinoma; Pancreatic cancer; Gastroesophageal junction adenocarcinoma	Carsgen Therapeutics, Ltd.
  AB-011		Monoclonal Antibody	Clinical Phase I	Solid tumors; Gastric cancer; Pancreatic adenocarcinoma
  ASKB-589	Claudin 18.2	Monoclonal Antibody	Clinical Phase II	Pancreatic cancer; Malignant solid tumors; Gastric cancer	Jiangsu Aosaikang Pharmaceutical Co., Ltd.
  Q-1802	Claudin 18.2; PD-L1	Bispecific Antibody	Clinical Phase I	Esophageal cancer; Pancreatic cancer; Advanced solid tumors; Gastric cancer	QureBio Ltd.
  MIL-93	Claudin 18.2	Monoclonal Antibody	Clinical Phase I	Gastric cancer; Pancreatic cancer; Advanced or metastatic solid tumors	Beijing Mabworks Biotech Co., Ltd.
  TST-001	Claudin 18.2; Complement cascade	Monoclonal Antibody	Clinical Phase IIa/Ib	Advanced gastric cancer; Advanced and/or metastatic solid tumors	Mabspace Biosciences (Suzhou) Co., Ltd.
  CMG-901	Claudin 18.2	ADC	Clinical Phase I	Advanced solid Tumors; Gastric cancer; Gastroesophageal junction adenocarcinoma; Pancreatic cancer	Keymed Biosciences Co.Ltd
  AMG-910	CD3; Claudin 18.2	Bispecific Antibody	Clinical Phase I	Gastric and gastroesophageal junction adenocarcinoma	Amgen
  M-108	Claudin 18.2	Monoclonal Antibody	Clinical Phase I	Advanced unresectable solid tumors	FutureGen Biopharmaceutical (Beijing) Co., Ltd
  LCAR-C18S	Claudin 18.2	CAR-T	Clinical Phase I	Gastric cancer	Nanjing Legend Biotech Co.
  SHR-A1904	Claudin 18.2	ADC	Clinical Phase I	Advanced solid cancer; Advanced pancreatic cancer	Shanghai Hengrui Pharmaceutical Co., Ltd
  ATG-022	Claudin 18.2	ADC	Preclinical	Solid tumors	Antengene Corp Ltd
  NBL-015	Claudin 18.2	Monoclonal Antibody	Preclinical	Pancreatic cancer Gastric cancer (esophagus-gastric junction carcinoma)	CSPC Pharmaceutical Group Co., Ltd.
  SYSA1801	Claudin 18.2	ADC	Clinical Phase I	Advanced solid tumors; Gastric cancer; Gastroesophageal junction cancer; Pancreatic cancer	CSPC ZhongQi Pharmaceutical Technology Co., Ltd.
  DR30303	Claudin 18.2	Monoclonal Antibody	Preclinical	Solid tumors	Doer Biologics
  SOT102	Claudin 18.2	ADC	Preclinical	Solid tumors	SOTIO Biotech/NBE-Therapeutics AG
  HX-s004	Claudin 18.2; PD-1	CAR-T	Preclinical	Gastric cancer	China Immunotech
  ZL-1211	Claudin 18.2	Monoclonal Antibody	Preclinical	Oncology	Zai Lab
  QL-1779	Claudin 18.2	Monoclonal Antibody	Preclinical	Advanced solid tumors	Qilu Pharmaceutical Co., Ltd.
  CTD101	Claudin 18.2	CAR-T	Preclinical	Gastric cancer	Nanjing Bioheng Biotech Co., Ltd.
  BA-1105	Claudin 18.2	Monoclonal Antibody	Preclinical	Esophageal cancer; Pancreatic cancer; Gastric cancer	Boan Biotech
  TJ-CD4B/ABL111	4-1BB; Claudin 18.2	Bispecific Antibody	Clinical Phase I	Advanced or metastatic solid tumors	ABL Bio/ I-Mab
  BNT141	CLDN18.2	Monoclonal Antibody	Clinical Phase I/II	Solid tumors	BioNTech SE
  GB264	CD3; Claudin 18.2	Bispecific Antibody	Preclinical	GI cancers	Genor Biopharma Co. Ltd.
  GB-7004	Claudin 18.2; Complement cascade	Monoclonal Antibody	Preclinical	Gastric cancer	Shanghai Genechem Co., LTD.
  HBM-1029	Claudin 18.2	Monoclonal Antibody	Preclinical	Solid tumors	Harbour BioMed
  LM302	Claudin 18.2	ADC	Preclinical	Gastrointestinal cancer	LaNova Medicines Ltd.
  RC-118	Claudin 18.2	ADC	Clinical Phase I	Solid tumors	RemeGen
  PT-886	CD47; Claudin 18.2	Bispecific Antibody	Preclinical	Pancreatic & gastric cancer	Phanes Therapeutics

  Data of table 1 comes from https://clinicaltrials.gov/ and https://www.pharnexcloud.com/

• References

  1. Hou, Jianghui; Konrad, Martin (2010-01-01), L. Yu, Alan S. (ed.), "Chapter 7 - Claudins and Renal Magnesium Handling", Current Topics in Membranes, Academic Press, 65, pp. 151–176, doi:10.1016/s1063-5823(10)65007-7, retrieved 2020-10-22

  2. Szaszi, Katalin; Amoozadeh, Yasaman (2014-01-01), Jeon, Kwang W. (ed.), "Chapter Six - New Insights into Functions, Regulation, and Pathological Roles of Tight Junctions in Kidney Tubular Epithelium", International Review of Cell and Molecular Biology, Academic Press, 308, pp. 205–271, doi:10.1016/b978-0-12-800097-7.00006-3, retrieved 2020-10-22

  3. Greene, Chris; Campbell, Matthew; Janigro, Damir (2019-01-01), Lonser, Russell R.; Sarntinoranont, Malisa; Bankiewicz, Krystof (eds.), "Chapter 1 - Fundamentals of Brain–Barrier Anatomy and Global Functions", Nervous System Drug Delivery, Academic Press, pp. 3–20, doi:10.1016/b978-0-12-813997-4.00001-3, ISBN 978-0-12-813997-4, retrieved 2020-10-22

  4. Morin PJ. Claudin proteins in human cancer: promising new targets for diagnosis and therapy. Cancer Res. 2005;65(21):9603–6.

  5. Sahin U, Koslowski M, Dhaene K, Usener D, Brandenburg G, Seitz G, Huber C, Tureci O. Claudin-18 splice variant 2 is a pan-cancer target suitable for therapeutic antibody development. Clin Cancer Res. 2008;14(23):7624–34.

  6. Sanada Y, Oue N, Mitani Y, Yoshida K, Nakayama H, Yasui W. Down-regulation of the claudin-18 gene, identified through serial analysis of gene expression data analysis, in gastric cancer with an intestinal phenotype. J Pathol. 2006;208(5):633–42.

  7. Fontana E, et al. Ther Adv Med Oncol. 2016;8:113-125.

  8. Yang B, et al. J Exp Clin Cancer Res. 2019;38:283.

  9. Zhu G, et al. Sci Rep. 2019;9:8420.

  10. Selim JH, Shaheen S, Sheu WC, Hsueh CT. Targeted and novel therapy in advanced gastric cancer. Exp Hematol Oncol. 2019; 8: 25.

  11. Singh, P., Toom, S. & Huang, Y. Anti-claudin 18.2 antibody as new targeted therapy for advanced gastric cancer. J Hematol Oncol 10, 105 (2017). https://doi.org/10.1186/s13045-017-0473-4

  12. Haishan Lin, Richard Zhang. Development of anti-human CLDN18.2 monoclonal antibody as cancer therapeutics. Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B73.

  13. Zhan, X.; Lin, B.; Li, Z.; Li, J.; Lin, H.; Chen, L.; Jiang, H.; Wu, M.; Xiao, J.; Peng, X.; et al. Phase I trial of Claudin 18.2-specific chimeric antigen receptor T cells for advanced gastric and pancreatic adenocarcinoma. J. Clin. Oncol. 2019, 37, 2509