The invention discloses a separation and purification method for a monoclonal antibody. The method includes the steps that cell culture supernatant containing the monoclonal antibody is subjected to Protein A affinity chromatography so as to obtain a first elution component; the first elution component is placed for a specified time at the room temperature with a specified pH value kept so as to obtain the first elution component subjected to low-pH-value inactivated virus treatment; the first elution component subjected to low-pH-value inactivated virus treatment is neutralized and then subjected to cation exchange chromatography so as to obtain a second elution component; the second elution component is diluted and then subjected to anion exchange chromatography so as to obtain a through flow; the through flow is filtered with a virus removal filter so as to obtain filter liquor; the filter liquor is subjected to ultrafiltration concentration, filter wash and buffer solution replacement in sequence so as to obtain stoste of the monoclonal antibody. The method is simple and easy to implement, and the endotoxin content and other quality indexes of the obtained product meet the quality standard of monoclonal antibody drugs.