TABLE 2:

Steps for immobilization of MDP compounds

Step	Biacore 3000	BI-4500
Mount the chip	“Dock”: for docking the chip “Prime”: for equilibrating the system with running buffer PBS at 5 μL/min	“Startup”: for cleaning the prism stage, mounting the chip, and purging the system with running buffer at 60 μL/min
Make a 1:1 EDC/NHS solution	“Dilute”: to mix NHS solution with EDC solution	“Add” and “Stir”: to add NHS solution to EDC-containing vial and mix well
Inject EDC/NHS to activate carboxylic acid groups to amine-reactive NHS-esters	“Inject”: pass EDC/NHS solution through the gold chip for 7 minutes	“Inject”: flow 200 μL of EDC/NHS solution over the chip
Wash with PBS to remove residual EDC/NHS	“Inject”: add PBS for 2 mins	“Inject”: to inject 200 μL of PBS
Coupling reaction between MDP containing an amino group and NHS-esters results in the formation of an amide bond.	“Inject”: flow MDP solution over the chip for 7 minutes	“Inject”: add 200 μL of MDP solution
Wash	“Inject”: wash with PBS for 2 minutes	“Inject”: to inject 200 μL of PBS
Deactivate unreacted carboxylic acid group with ethanolamine (EA)	“Inject”: flow 1M EA solution over the chip for 7 minutes	“Inject”: inject 200 μL of 1M EA
Wash with SDS	“Inject”: flow SDS buffer over the chip for 3 minutes	“Inject”: inject 200 μL of SDS buffer
Wash with PBS	“Unclog”: flow PBS buffer over the chip to wash away all residues at a high flow rate	“Inject”: inject 400 μL of PBS