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PerkinElmer Life & Analytical Sciences, Chalfont Road, Seer Green, Beaconsfield, Bucks. HP9 2FX. UK The widening scope of Proteomics and the increasing demands on high-throughput analysis has driven the need for instrumentation which can handle a wider range of samples with improved performance and ruggedness. A new MALDI-TOF instrument - equipped with both collisional cooling and orthogonal injection has been developed. This unique design approach effectively decouples ion production from time-of-flight measurements, making the instrument much more robust, much faster and far easier to use. Independence of operating conditions allows the system to be optimized for maximum ion yield and spectral resolution. At the same time, high mass accuracy is achieved and maintained without the need for internal calibration - a critical success factor for protein analysis, especially in a high-throughput environment. Decoupling the ionization and mass analysis processes permits more flexibility in the design of a sample target. Traditionally, samples are prepared on reusable metal targets where sample carryover is a concern. Single-use sample plates in micro-titer format, were developed to assure reproducible analysis without contamination. The new single-use plates provide convenient sample spot confinement and deliver performance equivalent to standard reusable plates. In this presentation, the use of orthogonal technology with MALDI-TOF will be reviewed. The performance of this new instrument will be demonstrated on proteins and protein digests prepared by gel- and LC-based separations. Performance criteria including mass accuracy, mass resolution, and sensitivity will be examined. Mass spectra of protein mixtures across a wide mass range, acquired using a single set of instrument parameters, will be presented to illustrate the wide mass range available. Mass accuracy in the low ppm range and sensitivity in the sub fmol range provide the opportunity to successfully analyze low quantities of sample.
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