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Construction of Anti-HER2 Recombinants as Targeting Modules for a Drug-delivery System Against HER2-positive Cells
- Source :
- Anticancer Research. 38:4319-4325
- Publication Year :
- 2018
- Publisher :
- Anticancer Research USA Inc., 2018.
-
Abstract
- Background/aim Recombinant antibodies have been investigated and used in applications such as targeting modules of drug-delivery systems (DDS) against cancers. This study aimed to prepare recombinant antibodies against HER2, containing sortase A (SrtA) recognition sequence, that are applicable as targeting modules in DDS after linkage with the drug-carrier containing oligoglycine-acceptor peptide by SrtA transpeptidation. Materials and methods The recombinant trastuzumab fragment antibodies (scFvs and Fab) with the SrtA-recognition motif (LPXTG) at their C-terminal were constructed and expressed in Escherichia coli and Chinese hamster ovary (CHO) cells, respectively. The reactivity of the purified recombinant antibodies towards HER2-expressing cells was also evaluated via immunofluorescent staining. Results Fab demonstrated higher yield and purity and better reactivity towards HER2-expressing cells (HCT-15 and HeLa) when compared to scFvs. Conclusion The CHO expression system possesses superior yield and purity when compared to the E. coli expression system with respect to the preparation of recombinant antibodies applicable in targeting modules for DDS (DDS-TM). Moreover, a Fab variant prepared in this study demonstrated the potential to be a DDS-TM against HER2-expressing cancer cells.
- Subjects :
- 0301 basic medicine
Cancer Research
Receptor, ErbB-2
CHO Cells
medicine.disease_cause
01 natural sciences
law.invention
Immunoglobulin Fab Fragments
03 medical and health sciences
Cricetulus
law
Cricetinae
medicine
Animals
Humans
Molecular Targeted Therapy
Escherichia coli
biology
Chemistry
Chinese hamster ovary cell
General Medicine
Trastuzumab
Molecular biology
Recombinant Proteins
0104 chemical sciences
010404 medicinal & biomolecular chemistry
030104 developmental biology
Oncology
Sortase A
Cancer cell
biology.protein
Recombinant DNA
Antibody
Single-Chain Antibodies
Subjects
Details
- ISSN :
- 17917530 and 02507005
- Volume :
- 38
- Database :
- OpenAIRE
- Journal :
- Anticancer Research
- Accession number :
- edsair.doi.dedup.....a0418ddc62836fb6e0f1c89e9c8675b5
- Full Text :
- https://doi.org/10.21873/anticanres.12731