These were cultured with Dulbeccos modified Eagles medium (D5796, Sigma-Aldrich, Saint-Louis, MO) supplemented with 10% heat-inactivated fetal bovine serum (Sigma-Aldrich), 100 U/mL penicillin, and 100 g/mL streptomycin (Nacalai Tesque)
Taking a different approach, we then engineered the fusion molecule to create steric hindrance around the peptide and block its degradation by proteases either by creating an N-linked glycosylation site at position 25 of the GLP-1 analogue peptide (Ab#2_GLP1 W25N/V27S) or by introducing a disulphide bridge between the exendin-4 peptide and the linker (Ab#2_DSB#1)
References: Andres, E., Loukili, N
Use a fresh, sterile syringe every time
Seek immediate medical attention for back pain with fever, bowel or bladder dysfunction, progressive leg weakness, severe abdominal pain, or saddle anesthesia
Adjusting dosage or titration speed can make a big difference