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Encyclopedia of Experiments: Immunology

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A Technique to Assess the In Vivo Localization of Tumor-Specific Antibodies

 

A Technique to Assess the In Vivo Localization of Tumor-Specific Antibodies

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Take a human ovarian cancer cell suspension. Add a basement membrane matrix medium, and keep on ice to avoid gel formation.

Load the mixture into a syringe and inject subcutaneously into an athymic nude mouse. Remove the needle once the matrix undergoes gelation, preventing mixture expulsion.

The lack of an immune response in the immunodeficient mouse preserves the viability of the injected cells. The gel's extracellular matrix proteins and growth factors provide growth-promoting signals, facilitating tumor formation.

Tumor cells overexpress surface-bound folate receptor alpha-1, or FOLR1.

Take FOLR1-specific antibodies. Add a fluorescent dye and incubate, allowing dye conjugation with the antibodies.

Intravenously inject the labeled antibodies into the anesthetized tumor-bearing mouse.

Antibodies reach the tumor via circulation and bind to FOLR1 on tumor cells, fluorescently labeling the cell surface.

Perform in vivo fluorescence imaging. Upon excitation, the dye emits fluorescence, facilitating the assessment of antibody localization in the tumor.

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