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In JoVE (1)
Other Publications (5)
Articles by Michael Lerch in JoVE
JoVE General
A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
1Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco - UCSF, 2Buck Institute for Age Research, 3Department of Chemistry, Purdue University
Lectin-conjugated POROS beads were employed for HPLC. Glycopeptide standards served as positive and negative controls. MARS-14 depleted, trypsin-digested human plasma was chromatographed and flow-through (FT) and bound fractions collected for ESI-LC-MS/MS analyses. Glycopeptides were enriched in the bound fraction as compared to FT.
Other articles by Michael Lerch on PubMed
Verification of the Plan Dosimetry for High Dose Rate Brachytherapy Using Metal-oxide-semiconductor Field Effect Transistor Detectors
The feasibility of a recently designed metal-oxide-semiconductor field effect transistor (MOSFET) dosimetry system for dose verification of high dose rate (HDR) brachytherapy treatment planning was investigated. MOSFET detectors were calibrated with a 0.6 cm3 NE-2571 Farmer-type ionization chamber in water. Key characteristics of the MOSFET detectors, such as the energy dependence, that will affect phantom measurements with HDR 192Ir sources were measured. The MOS-FET detector was then applied to verify the dosimetric accuracy of HDR brachytherapy treatments in a custom-made water phantom. Three MOSFET detectors were calibrated independently, with the calibration factors ranging from 0.187 to 0.215 cGy/mV. A distance dependent energy response was observed, significant within 2 cm from the source. The new MOSFET detector has a good reproducibility (<3%), small angular effect (<2%), and good dose linearity (R2=1). It was observed that the MOSFET detectors had a linear response to dose until the threshold voltage reached approximately 24 V for 192Ir source measurements. Further comparison of phantom measurements using MOSFET detectors with dose calculations by a commercial treatment planning system for computed tomography-based brachytherapy treatment plans showed that the mean relative deviation was 2.2 +/- 0.2% for dose points 1 cm away from the source and 2.0 +/- 0.1% for dose points located 2 cm away. The percentage deviations between the measured doses and the planned doses were below 5% for all the measurements. The MOSFET detector, with its advantages of small physical size and ease of use, is a reliable tool for quality assurance of HDR brachytherapy. The phantom verification method described here is universal and can be applied to other HDR brachytherapy treatments.
In Vivo Verification of Superficial Dose for Head and Neck Treatments Using Intensity-modulated Techniques
Skin dose is one of the key issues for clinical dosimetry in radiation therapy. Currently planning computer systems are unable to accurately predict dose in the buildup region, leaving ambiguity as to the dose levels actually received by the patient's skin during radiotherapy. This is one of the prime reasons why in vivo measurements are necessary to estimate the dose in the buildup region. A newly developed metal-oxide-semiconductor-field-effect-transistor (MOSFET) detector designed specifically for dose measurements in rapidly changing dose gradients was introduced for accurate in vivo skin dosimetry. The feasibility of this detector for skin dose measurements was verified in comparison with plane parallel ionization chamber and radiochromic films. The accuracy of a commercial treatment planning system (TPS) in skin dose calculations for intensity-modulated radiation therapy treatment of nasopharyngeal carcinoma was evaluated using MOSFET detectors in an anthropomorphic phantom as well as on the patients. Results show that this newly developed MOSFET detector can provide a minimal but highly reproducible intrinsic buildup of 7 mg cm(-2) corresponding to the requirements of personal surface dose equivalent Hp (0.07). The reproducibility of the MOSFET response, in high sensitivity mode, is found to be better than 2% at the phantom surface for the doses normally delivered to the patients. The MOSFET detector agrees well with the Attix chamber and the EBT Gafchromic film in terms of surface and buildup region dose measurements, even for oblique incident beams. While the dose difference between MOSFET measurements and TPS calculations is within measurement uncertainty for the depths equal to or greater than 0.5 cm, an overestimation of up to 8.5% was found for the surface dose calculations in the anthropomorphic phantom study. In vivo skin dose measurements reveal that the dose difference between the MOSFET results and the TPS calculations was on average -7.2%, ranging from -4.3% to -9.2%. The newly designed MOSFET detector encapsulated into a thin water protective film has a minimal reproducible intrinsic buildup recommended for skin dosimetry. This feature makes it very suitable for routine IMRT QA and accurate in vivo skin dosimetry.
In Vivo Real-time Rectal Wall Dosimetry for Prostate Radiotherapy
Rectal balloons are used in external beam prostate radiotherapy to provide reproducible anatomy and rectal dose reductions. This is an investigation into the combination of a MOSFET radiation detector with a rectal balloon for realtime in vivo rectal wall dosimetry. The MOSFET used in the study is a radiation detector that provides a water equivalent depth of measurement of 70 microm. Two MOSFETs were combined in a face-to-face orientation. The reproducibility, sensitivity and angular dependence were measured for the dual MOSFET in a 6 MV photon beam. The dual MOSFET was combined with a rectal balloon and irradiated with hypothetical prostate treatments in a phantom. The anterior rectal wall dose was measured in real time and compared with the planning system calculated dose. The dual MOSFET showed angular dependence within +/-2.5% in the azimuth and +2.5%/-4% in the polar axes. When compared with an ion chamber measurement in a phantom, the dual MOSFET agreed within 2.5% for a range of radiation path lengths and incident angles. The dual MOSFET had reproducible sensitivity for fraction sizes of 2-10 Gy. For the hypothetical prostate treatments the measured anterior rectal wall dose was 2.6 and 3.2% lower than the calculated dose for 3DCRT and IMRT plans. This was expected due to limitations of the dose calculation method used at the balloon cavity interface. A dual MOSFET combined with a commercial rectal balloon was shown to provide reproducible measurements of the anterior rectal wall dose in real time. The measured anterior rectal wall dose agreed with the expected dose from the treatment plan for 3DCRT and IMRT plans. The dual MOSFET could be read out in real time during the irradiation, providing the capability for real-time dose monitoring of the rectal wall dose during treatment.
A Lectin Affinity Workflow Targeting Glycosite-specific, Cancer-related Carbohydrate Structures in Trypsin-digested Human Plasma
Glycans are cell-type-specific, posttranslational protein modifications that are modulated during developmental and disease processes. As such, glycoproteins are attractive biomarker candidates. Here, we describe a mass spectrometry-based workflow that incorporates lectin affinity chromatography to enrich for proteins that carry specific glycan structures. As increases in sialylation and fucosylation are prominent among cancer-associated modifications, we focused on Sambucus nigra agglutinin (SNA) and Aleuria aurantia lectin (AAL), lectins which bind sialic acid- and fucose-containing structures, respectively. Fucosylated and sialylated glycopeptides from human lactoferrin served as positive controls, and high-mannose structures from yeast invertase served as negative controls. The standards were spiked into Multiple Affinity Removal System (MARS) 14-depleted, trypsin-digested human plasma from healthy donors. Samples were loaded onto lectin columns, separated by HPLC into flow-through and bound fractions, and treated with peptide: N-glycosidase F to remove N-linked glycans. The deglycosylated peptide fractions were interrogated by ESI HPLC-MS/MS. We identified a total of 122 human plasma glycoproteins containing 247 unique glycosites. Importantly, several of the observed glycoproteins (e.g., cadherin 5 and neutrophil gelatinase-associated lipocalin) typically circulate in plasma at low nanogram per milliliter levels. Together, these results provide mass spectrometry-based evidence of the utility of incorporating lectin-separation platforms into cancer biomarker discovery pipelines.
Real-time in Vivo Dosimetry with MOSFET Detectors in Serial Tomotherapy for Head and Neck Cancer Patients
A real-time dose verification method using a recently designed metal oxide semiconductor field effect transistor (MOSFET) dosimetry system was evaluated for quality assurance (QA) of intensity-modulated radiation therapy (IMRT).
