SEQUENCE AND STRUCTURE OF V-H DOMAIN FROM NATURALLY-OCCURRING CAMEL HEAVY-CHAIN IMMUNOGLOBULINS LACKING LIGHT-CHAINS

We cloned 17 different PCR fragments encoding VH genes of camel (Camelus dromedarius). These clones were derived from the camel heavy chain immunoglobulins lacking the light chain counterpart of normal immunoglobulins. Insight into the camel V-H sequences and structure may help the development of single domain antibodies. The most remarkable difference in the camel V-H, consistent with the absence of the V-L interaction, is the substitution of the conserved Leu45 by an Arg or Cys. Another noteworthy substitution is the Leu11 to Ser. This amino acid normally interacts with the C(H)1 domain, a domain missing in the camel heavy chain immunoglobulins. The nature of these substitutions agrees with the increased solubility behavior of an isolated camel V-H domain. The V-H domains of the camels are also characterized by a long CDR3, possibly compensating for the absence of the V-L contacts with the antigen. The CDR3 lacks the salt bridge between Arg94 and Asp101. However, the frequent occurrence of additional Cys residues in both the CDR1 and CDR3 might lead to the formation of a second internal disulfide bridge, thereby stabilizing the CDR structure as in the DAW antibody. Within CDRs of the camel V-H domains we observe a broad size distribution and a different amino acid pattern compared with the mouse or human V-H. Therefore the camel hypervariable regions might adopt structures which differ substantially from the known canonical structures, thereby increasing the repertoire of the camel antigen binding sites within a V-H.