Development of a novel ELISA for human insulin using monoclonal antibodies produced in serum-free cell culture medium.

Objectives: Development of an ELISA for human insulin that utilizes monoclonal antibodies (mAbs) produced in serum-free medium.

Design And Methods: Insulin mAbs were produced in vitro by hybridomas in serum-free medium. A two-step ELISA was developed to replace bovine insulin (standard) and bovine serum albumin (assay buffer) with non-animal reagents.

Serum-free hybridoma culture: ethical, scientific and safety considerations.

Despite considerable progress in the development of cell culture techniques, including the development of the serum- and protein-free media that now routinely support hybridoma and mammalian cell growth, fetal bovine serum (FBS) supplemented media are still commonly used: a practice that raises ethical, scientific and safety concerns. The use of FBS in hybridoma culture media is examined here, with regards to the development and production of monoclonal antibodies (mAbs), and it is our recommendation that researchers adopt serum-free cell culture methods to reduce animal use in this area.