An In Vivo Immunofluorescence Localization Method to Study Antibody Biodistribution

Published: January 31, 2024

Abstract

Source: Wischhusen, J. C., et al. In Vivo Immunofluorescence Localization for Assessment of Therapeutic and Diagnostic Antibody Biodistribution in Cancer Research. J. Vis. Exp. (2019).

This video demonstrates an in vivo immunofluorescence localization (IVIL) technique to study the biodistribution of tumor-targeting antibodies in a murine model of breast cancer. Upon the intravenous injection of a tumor-specific antibody in the mouse, the tumor is surgically isolated, and the localization of the antibody within the tumor tissue is confirmed via ex vivo immunostaining.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Transgenic mouse model of breast cancer development Observe mice from the desired cancer model for the appropriate tumor growth via palpation or caliper measurement before proceeding. NOTE: Here, the transgenic murine model of breast cancer development (FVB/N-Tg(MMTV-PyMT)634Mul/J) (MMTV…

Divulgations

The authors have nothing to disclose.

Materials

Animal Model
FVB/N-Tg(MMTV-PyMT)634Mul/J The Jackson Laboratory 2374 Females, 4-6 weeks of age
Animal Handling Supplies
27G Catheter VisualSonics Please call to order Vevo MicroMarker Tail Vein Access Cannulation Kit
Alcohol Wipes Fisher Scientific 22-246073
Gauze Sponges (4" x 4" 16 Ply) Cardinal Health 2913
Heat Lamp Morganville Scientific HL0100
Isoflurane Henry Schein Animal Health 29404
Ophthalmic Ointment Fisher Scientific NC0490117
Surgical Tape 3M 1530-1
Tissue Collection
Disposable Base Molds Fisher Scientific 22-363-556
Optimal Cutting Temperature (OCT) Medium Fisher Scientific 23-730-571
Surgical London Forceps Fine Science Tools 11080-02
Surgical Scissors Fine Science Tools 14084-08
Antibodies
AlexaFluor-488 goat anti-rat IgG Life Technologies A-11006
AlexaFluor-546 goat anti-rabbit IgG Life Technologies A-11010
AlexaFluor-594 goat anti-human IgG Life Technologies A11014
Human IgG Isotype Control Novus Biologicals NBP1-97043
Humanized anti-netrin-1 antibody Netris Pharma contact@netrispharma.com
Rabbit anti-Mouse CD276 (B7-H3) Abcam ab134161 EPNCIR122 Clone
Rat anti-Mouse CD31 BD Biosciences 550274 MEC 13.3 Clone
Reagents
Bovine Serum Albumin (BSA) Sigma-Aldrich A2153-50G
Clear Nail Polish Any local drug store
Indocyanine Green – NHS Intrace Medical ICG-NHS ester
Mounting Medium ThermoFisher Scientific TA-006-FM
Normal Goat Serum Fisher Scientific ICN19135680
Paraformaldehyde (PFA) Fisher Scientific AAJ19943K2
Sterile Phosphate Buffered Saline (PBS) ThermoFisher Scientific 14190250
Triton-X 100 Sigma-Aldrich T8787
Supplies
Adhesion Glass Slides VWR 48311-703
Desalting Columns Fisher Scientific 45-000-148
Glass Cover Slips Fisher Scientific 12-544G
Hydrophobic Barrier Pen Ted Pella 22311
Microcentrifuge Tubes Fisher Scientific 05-402-25
Slide Staining Tray VWR 87000-136
Software
FIJI LOCI, UW-Madison. Version 4.0 https://fiji.sc/

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Citer Cet Article
An In Vivo Immunofluorescence Localization Method to Study Antibody Biodistribution. J. Vis. Exp. (Pending Publication), e21891, doi: (2024).

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