Antibody-based assay for galacturonan deesterification on wood pulp fibers during bleaching

Antibodies recognizing galacturonan were used in immunoassays for industrial unbleached and hydrogen peroxide-bleached chemithermomechanical pulp (CTMP). The assays were based on a colorimetric method using specific monoclonal antibody recognition of acidic and methylated homogalacturonan, respectively, on the pulp fiber surfaces. Alkaline phosphatase-conjugated antibodies were linked to the galacturonan specific antibodies, and an alkaline phosphatase substrate (p-nitrophenylphosphate) was used to develop a yellow reaction product that can be followed spectrophotometrically. Together the newly developed immunoassays were able to describe the deesterification, specifically, of surface-localized galacturonan on CTMP, induced by alkaline hydrogen peroxide bleaching. Unbleached CTMP showed relatively more labeling by methylated galacturonan recognizing antibodies, whereas bleached CTMP was relatively more labeled for acidic galacturonan. The increase in negative charge of the bleached pulp was also shown by polyelectrolyte titration; the negative surface charge was 9.3 μeq g⁻¹ in unbleached CTMP and 21.7 μeq g⁻¹ in bleached pulp.