NR ATKU
AU Hartman,K.P.; Curin Serbec,V.
TI Properties of the PrPsc - specific mAb V5B2 guarantee a useful diagnostic tool
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-32
PT Konferenz-Poster
AB
Prion diseases are neurodegenerative lethal diseases that affect humans and animals. Protein infective particle, PrPsc, a conformational isoform of the normal prion protein, PrPc, is thought to be a causative agent. For research and for diagnostics it is very important to set forth a new specific tool for identification of PrPsc but not its normal counterpart PrPc. The antibodies currently used to detect the infective agent namely do not differentiate between PrPsc and PrPc.
We have already reported on a PrPsc-specific mAb V5B2 (Curin Serbec et al., 2004), which was also proven to be useful in post mortem test without proteinase K treatment. Here we present the biological properties of the hybridoma cell line V5B2 and its clone B2B2.
Both cell strains had steep growth curve in logarithm phase and population doubling time of around 8 hours. Other growth properties, important for in vitro cell culturing, were also adequate. In any case, the production of antibodies exceeded 100 µg/ml. mAbs could be easily stored as supernatants at +4 ° or at -20 °, they could be purified and stored in saline at -20 ° or could be lyophilised. Stability of mAbs was estimated after four years. Supernatants, kept at -20 °, retained full activity, whereas lyophilised product's reactivity dropped 5 times.
We show that V5B2 is a very stable mAb and therefore suitable for production in large quantities. Since it is an IgG, it can be readily used in different immunoassays in research or in diagnostic tests.
AD Katrina Pretnar Hartman, Blood Transfusion Centre of Slovenia, Slovenia; Vladka Curin Serbec, Blood Transfusion Centre of Slovenia, Slovenia
SP englisch
PO Deutschland