UV ([lambda]= 254 nm, 6 W) was irradiated to EPC solution in DMSO and HmPVA--EPC conjugate solution
in distilled water.
The results for EPX indicated that the amount of mAb deposited on the membrane should be 250 ng/strip, and the amount of mAb in the conjugate solution
should be 0.4 [micro]g/strip to avoid excessively high signal in the detector and to optimize sensitivity.
Working wash solution and working conjugate solution
were prepared separately.
(21) Finally it was immersed in antihuman IgG-IGNPs conjugate solution
at 37[degrees]C for 30 min, and then rinsed by 0.02 mol/L PBS (pH 7.4) to remove unbound antihuman IgG-IGNPs conjugate.
For the study, 100 mL of magnetic microspheres at 4000 [micro]L and 50 [micro]L of TSH calibrator were incubated for 20 min, and the microspheres were washed and transferred to 50 [micro]L of the conjugate solution
. After incubation for 5 min, the microspheres were washed and transferred to 50 [micro]L of Pretrigger solution.
To the above conjugate solution
, an equal volume of ethylene glycol was added and kept at -30 [degrees]C in aliquots for future use.
The hydrodynamic radius (Rh) was estimated by the dynamic light scattering (DLS) of aqueous conjugate solutions
(0.5 wt%) in phosphate buffer (pH 7.4) at 22[degrees] C at a scattering angle of 173[degrees] on Zetasizer Nano-ZS (Malvern, UK).
To 250 [micro]L of the diluted conjugate solutions
, 250 [micro]L of 0.5 M phosphate buffer pH 8.0 and 500 [micro]L of Ellman's reagent were added.