Structural features of a murine monoclonal anti-ssDNA autoantibody

RODGER G SMITH, DEAN W BALLARD, PETER R BLIER, PATTY E PACE, ALFRED L M BOTHWELL, JAMES N HERRON, ALLEN B EDMUNDSON, EDWARD W VOSS JR

Abstract


Binding properties ofa murine (NZB x NZWF 1) monoclonal anti-ssDNA autoantibody (BV04-01) was compared with another anti-ssDNA autoantibody mV16-19). Both monoclonal antibodies preferentially bound ssDNA and poly(dT) relative to poly(dU). Sigmficant binding of synthetic polynucleotides was detectable when the homopolymer exceeded 5 pyrimidines in length. Binding showed an inverse temperature dependence between 2 and 4O C, while variations in ionic strength showed minimal effects. The  primary structurc of the variable region of the heavy (IgG2b) and light (kappa) chains comprising BV04-01 was determined using primer extension methodology. A polynucleotide synthesized to correspond wit11 unique sequences within heavy chain CDR2 was used as a gene probe in Northern blot analyses.

Keywords


Binding properties; polynucleotide; gene probe; homopolymer; autoimmune diseases; monoclonal antibodies.

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