The invention discloses a method for sorting and screening nano
antibody through
flow cytometry. The method comprises the following steps: (1) collecting
peripheral blood B
lymphocyte of a camelidae animal with immune
target antigen; (2) sorting the B
lymphocyte as a single
cell by applying
target antigen through the
flow cytometry; (3) directly performing reverse transcriptional reaction on the sorted single B
lymphocyte to generate cDNA; (4) taking the cDNA as a template, performing PCR amplification on an
antibody heavy chain sequence and recycling an amplified product; (5) taking the amplified product in the step (4) as a template, and performing PCR amplification on CH1 and CH2 sequence coding genes of the
antibody; (6) taking the amplified product in the step (4) which is amplified to be negative in the step (5) as a template, and performing PCR amplification on a VHH segment coding
gene of the antibody; (7)
cloning a VHH segment obtained in the step (6) into an
expression vector, and expressing the VHH segment in host
bacteria; and (8) identifying the nano antibody which is pressed in the step (7). The method disclosed by the invention is simple to operate, is short in screening period, avoids phage
pollution risk, improves screening efficiency, reduces
workload greatly, is low in economic cost, and is suitable for industrial large-batch screening requirements.