Study of pilling assessment is very much essential to specify the performance and serviceability of fabrics. This paper describes the suitable and standard method of pilling assessment of cotton knitted fabrics. Especially for cotton fabrics, pilling and hairiness were not a problem in earlier period but recently those have become a serious problems in apparel sector. The pilling resistance of fabrics depends on some important parameters like fiber type, yarn spinning technique, fabric type and finishing process.this study, four types of cotton knit fabrics (single jersey, 1 × 1 rib, plain interlock & pique) have been used. Bio-polishing (an enzymatic treatment) was applied on selected cotton fabrics after scouring-bleaching and before dyeing and the effect of this on pilling resistance and weight loss of the fabrics were observed. Studies on this field were not carried out previous time in large scale, and hence it was not possible to observe the facts clearly. This investigation was specialy carried out under universal operational conditions, which are established in industry.