Journal
/
Cancer Research
/
Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo
JoVE Journal
Cancer Research
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Cancer Research
Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo

Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo

DOI:
10.3791/62018-v

•

10:35 min

•

February 05, 2021

•

, , , , , , ,
1Immune Regulation Group,Helmholtz Centre for Infection Research, 2Infection Immunology Group, Institute of Medical Microbiology, Infection Prevention and Control, Health Campus Immunology, Infectiology and Inflammation,Otto-von-Guericke University Magdeburg, 3Institute for Molecular and Clinical Immunology, Health Campus Immunology, Infectiology and Inflammation,Otto-von-Guericke University Magdeburg, 4Cellular Proteome Research,Helmholtz Centre for Infection Research, 5Experimental Pneumology, University Hospital of Pneumology, Health Campus Immunology, Infectiology and Inflammation,Otto-von-Guericke University Magdeburg

Chapters

  • 00:04Introduction
  • 00:50Anti-DEC-205 (CD205) Production
  • 02:47Anti-DEC-205 Antibody Purification
  • 04:40Chemical Conjugation
  • 06:43Chemical Conjugation Verification
  • 08:22Results: Representative Purified anti-DEC-205 Antibody Conjugation Characterization and Analysis
  • 09:52Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a protocol for the chemical conjugation of the model antigen ovalbumin to an endocytosis receptor-specific antibody for in vivo dendritic cell targeting. The protocol includes purification of the antibody, chemical conjugation of the antigen, as well as purification of the conjugate and the verification of efficient conjugation.

Tags

Keywords: DEC-205 AntibodyOVA AntigenChemical ConjugationProtein AntigenIn VitroIn VivoAnti-tumor VaccinationNLDC-145 CellsCell CultureProtein G SepharoseAntibody Purification

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/62018?article_type=v

In Vitro Imaging and Quantification of the Drug Targeting Efficiency of Fluorescently Labeled GnRH Analogues
check_url/62018?article_type=v

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure
check_url/62018?article_type=v

Counting Proteins in Single Cells with Addressable Droplet Microarrays
check_url/62018?article_type=v

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol
check_url/62018?article_type=v

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment
check_url/62018?article_type=v

Isolation Protocol of Mouse Monocyte-derived Dendritic Cells and Their Subsequent In Vitro Activation with Tumor Immune Complexes
check_url/62018?article_type=v

A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
check_url/62018?article_type=v

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
check_url/62018?article_type=v

In Vivo Immunofluorescence Localization for Assessment of Therapeutic and Diagnostic Antibody Biodistribution in Cancer Research
check_url/62018?article_type=v

In Vivo Targeting of Xenografted Human Cancer Cells with Functionalized Fluorescent Silica Nanoparticles in Zebrafish

Read Article