This research work tries to achieve a multifunctional finishing on the cotton fabric by using environmentally friendly chemicals. Chitosan and N-(2-hydroxy) propyl-3-trimethyl ammonium chitosan chloride (HTCC) with different crosslinking agents including citric acid (CA), butane tetra carboxylic acid (BTCA), and glutaraldehyde (GA) were coapplied on the cotton fabric. This improves the laundering durability of antimicrobial treatment by formation of covalent bond between the crosslinking agent, antimicrobial agent, and cellulosic chains. Furthermore, it helps to enhance the crease resistance properties of the treated samples. The antimicrobial activities of the treated samples were tested after 15 laundering cycles. The whiteness and wrinkle recovery angle of the treated fabric were also examined. The glutaraldehyde treatment was superior to CA treatment in terms of prolonged antimicrobial activity of the samples after successive laundering. Also, the treated samples with glutaraldehyde showed a higher wrinkle recovery angle with excessive deterioration in whiteness when compared with CA and BTCA. The treated samples with CA and BTCA show good antimicrobial activity after successive laundering and improved wrinkle recovery angle. However, BTCA indicates a higher durability in comparison with CA and same durability after repeated laundering with glutaraldehyde. This can be achieved without yellowing and unpleasant odor.