the plate for binding to the soluble hgf flag indicating that these three mabs have very similar affinity a key biological activity of hgf is the ability to bind to its receptor cmet so the ability of the hul2g7 chl2g7 and l2g7 mabs to inhibit binding of hgf to met was compared met was used in the form of met fc and hgf in the form of hgf flag which were prepared as described patent application u s ser no 10 917 915 filed aug 13 2004 and kim et al clin cancer res 12 1292 2006 a microtiter plate was coated with 50 u03bcl well of 2 u03bcg ml each of two anti met mabs galaxy biotech in pbs overnight at 4 u00b0 c alternatively 2 u03bcg ml g higg fc may be used and blocked with 2 bsa for 1 hr at rt after washing the plates 50 u03bcl well of met fc 1 u03bcg ml was added to each well for 1 hr at rt followed after washing by 50 u03bcl well of human igg 10 u03bcg ml for 1 hr to reduce background after washing the plates 50 u03bcl well of hgf flag 0 5 u03bcg ml preincubated with various concentrations of mabs was added to each well for 1 hr after washing the plates were incubated with 50 u03bcl well of hrp m2 anti flag mab invitrogen and the bound hrp anti flag m2 detected by the addition of the substrate as described above fig5 shows that hul2g7 chl2g7 and l2g7 blocked binding of hgf to met equally well so these mabs have very.