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In the bio-polishing process, bio-degradable cellulase is used to attack the hair like cellulose substrate of fabric surface and it's action requires suitable and appropriate process conditions. For this reason, it is very much essential to know about the suitable process parameters like temperature, time and pH upon which the optimum effects of bio-polishing depend. This paper will be very supportive for the industrial people to select an appropriate time, temperature and pH for the bio-polishing of cotton fabrics with cellulase. In this research, bio-polishing was carried out with the cellulase at various process conditions and after that Ends/inch, Picks/inch, GSM, Pilling resistance and Whiteness of cotton fabrics have been determined. From this determination it was possible to assess the effects of bio-polishing.