Enrofloxacin is fluoroquinolone antibiotic which prohibited approved for treatment in animals. However, their residues in animal products can pose adverse side effects to consumer. Therefore, the maximum residue limit of these drugs has been enforced in many countries. In this study, the cDNA encoding VH and VL genes was amplified from monoclonal antibody which was specific to enrofloxacin clone 48, cloned and sequenced. The obtained sequences were compared in the NCBI databases by using blastp program. The results found that VH nucleotide was composed of about 399 bps and theirs deduced amino acids showed 80-85% degree identities to the Ig superfamily group. A totally 356 bps of VL nucleotide was found and showed the degree of identities of 97-100% with an immunoglobulin kappa light chain. Moreover, the CDR I, CDR II and CDR III of the VH and VL sequences were specified. The results indicated that the highest degree of VH sequence homology (highest relationship) found in accession no. B26471 with 80% homology. However, 100% of VL sequences were found similarly to accession no. AHJ10945.1 than the others. The obtained results provided the useful and important information for the further recombinant antibody construction and production against enrofloxacin antibiotic.