Efficient recombinant of Lym-1 scFv gene using multiple doubly- restricted DNA fragments

Xu Bao Shi, Paul H. Gumerlock, Linda Kroger, Gerald L Denardo, Sally J. DeNardo

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

In order to improve radioimmunotherapy of lymphoma, a Lym-1 single- chain antigen-binding (scFv) protein molecule was produced. Because the commonly used polymerase chain reaction (PCR) method frequently causes unexpected mutations, we developed a non-PCR method for scFv gene assembly. The method involved a stepwise of doubly-restricted DNA fragments and re- digestion of the resultant concatamers. Using this strategy, the Lym-1 scFv expression gene was readily constructed without mutations. The recombinant gene was cloned into an expression vector and scFv protein was expressed. The method can be used for other genes or DNA recombination.

Original languageEnglish (US)
Pages (from-to)139-143
Number of pages5
JournalCancer Biotherapy and Radiopharmaceuticals
Volume14
Issue number2
StatePublished - 1999

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ASJC Scopus subject areas

  • Cancer Research
  • Pharmacology
  • Oncology

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