Detection of sulfur mustard adducts in human callus by phage antibodies.

As part of a research program to develop novel methods for diagnosis of sulfur mustard exposure in the human skin the suitability of phage display was explored. Phage display is a relative new method that enables researchers to quickly evaluate a huge range of potentially useful antibodies, thereby bypassing the more costly and time-consuming hybridoma technique. The Tomlinson I and J phage libraries were used to select phage antibodies exhibiting affinity for sulfur mustard adducts on keratins, isolated from human callus.

Standard operating procedure for immunuslotblot assay for analysis of DNA/sulfur mustard adducts in human blood and skin.

A standard operating procedure has been developed for an immunoslotblot assay of sulfur mustard adducts to DNA in human blood and skin for use in a field laboratory. A minimum detectable level of exposure of human blood in vitro (> or = 50 nM) sulfur mustard is feasible with the assay. In the case of human skin, a 1 s exposure to saturated sulfur mustard vapor (830 mg/m(-3)) could still be detected.

Detection of single-strand breaks and base damage in DNA of human white blood cells as a tool for biological dosimetry of exposure to ionizing radiation.

An immunochemical assay has been adapted to detect DNA damage in whole blood at biologically relevant doses of ionizing radiation. Upon alkaline treatment of whole blood, both strand breaks and base damage (which is converted into strand breaks by the addition of damage-specific enzymes) are detected by using antibodies that specifically bind to single-strand DNA. Single-strand breaks can be detected immediately after irradiation at doses as low as 0.

Overview of low-level radiation exposure assessment: biodosimetry.

The capability to make diagnostic assessments of radiation exposure is needed to support triage of radiation casualties and medical treatment decisions in military operations. At the International Conference on Low-Level Radiation Injury and Medical Countermeasures session on biodosimetry in the military, participants reviewed the field of biomarkers, covering a wide range of biological endpoints. Participants evaluated early changes associated with exposure to ionizing radiation, including chromosomal and DNA damage, gene expression and associated proteins, and DNA mutations.

Immunochemical detection of sulfur mustard adducts with keratins in the stratum corneum of human skin.

As part of a program to develop methods for diagnosis of exposure to chemical warfare agents, we developed immunochemical methods for detection of adducts of sulfur mustard to keratin in human skin. Three partial sequences of keratins containing glutamine or asparagine adducted with a 2-hydroxyethylthioethyl group at the omega-amide function were synthesized and used as antigens for raising antibodies. After immunization, monoclonal antibodies were obtained with affinity for keratin isolated from human callus exposed to 50 microM sulfur mustard.