A　rabbit　phage　display　library　of　single　chain　Fv　(scFv)　antibodies　was　constructed　to　screen　specific　scFv　antibodies　against　melamine.　Rabbits　were　immunized　with　melamine　immunogens　prepared　via　glutaraldehyde　method.　Two　recombinant　scFv　antibodies　with　high　specificity　for　melamine,　termed　A9　and　B4,　were　isolated　via　library　panning.　SDS-PAGE　analysis　showed　that　the　soluble　A9-scFv　and　B4-scFv　antibodies　were　expressed　successfully　and　the　scFvs　showed　good　sensitivity　with　an　IC　50　of　12.　64　and　10.　32　μM　to　melamine,　respectively.　The　cross-reactivity　of　2　scFv　antibodies　with　cyanuric　acid　and　chloromycetin　were　below　1%.　The　high　similarity　of　2　scFv　nucleotide　sequences　indicated　that　they　may　originate　from　1　B　cell　clone　that　had　undergone　somatic　diversification.　As　a　consequence,　the　conjugation　synthesis　was　successful　and　the　selected　scFvs　could　be　used　for　sensitive　detection　of　melamine　in　foods　and　feeds　by　immunoassay.　©　2011　The　Korean　Society　of　Food　Science　and　Technology　and　Springer　Netherlands.