PRODUCTION OF A HUMAN RECOMBINANT ANTIBODY AGAINST SEROTYPE A CANDIDA ALBICANS

After using 3 different generations of antibody including human and non-humanhyperimmune sera, monoclonal antibodies and chimeric antibodies, more recently a newer approachhas been developed in which the antibody genes are cloned directly from a patient peripheral blymphocytesand expressed in a host like e. coli. in this study the candida albicans serotype a (nctc3135) mannan was purified using a modified fehling method and used for selection of humanrecombinant antibody (hrab) from a c. albicans phage antibody library. after four rounds of affinityselecting (panning), 2 predominant clones were chosen by dna fingerprinting and elisa. a 248amino acid dna fragment coding for anti-c. albicans mannan scfv was sequenced and cloned in apbad topo cloning vector to produce a soluble and phage free antibody. the analysis of antibodysequences by v base index (dnaplot) confirmed the human antibody origin with the vh4 family inv segment of heavy variable chain and vl3 (lambda 3) in j segment of the light variable chain. thisantibody fragment was purified using immobilised metal affinity chromatography (imac) andinmmunoblotted as a 31kda recombinant protein.