Pressure Cyclying Technology

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PCT (Pressure Cycling Technology), a rapid sample pre-treatment method for micro-scale number of samples, is of high robustness. It is a patented technology platform based on repeated cycles of hydrostatic pressure between ambient (14.69 PSI) and ultra-high (45,000 PSI) levels. These rapid changes in pressure are used to control biomolecular interactions for applications such as accelerated proteolytic digestion, and improved extraction of cellular components, including proteins and lipids.

The PCT-HD Sample Preparation System is a proven PCT-based workflow for processing small tissue samples for proteomic and other applications. PCT-HD combines efficient, hands-off tissue homogenization and protein extraction with pressure-enhanced digestion, for rapid, efficient and reproducible generation of peptides for MS analysis. This unique workflow combines two of PBI’s innovative sample preparation tools: the Barocycler 2320EXT and our patented PCT MicroPestles. PCT-HD and the Barocycler 2320EXT can provide unprecedented speed and reproducibility for biomarker discovery, yielding peptides ready for clean-up and MS within 2-4 hours from the start of tissue processing.

The Barocycler 2320EXT is a compact, bench top instrument that can process up to 16 samples simultaneously using PCT MicroTubes. Features that come standard with the 2320EXT include: data input and output options to accommodate validation and quality control; computer-operated control with touch screen programming and automatic data logging; and the ability to control multiple pressure cycling parameters, such as the rate of pressure increase/decrease, both the high and low pressure levels, and the shape of the pressure profile (e.g., sine wave, square wave, and others). In addition, the 2320EXT is available with a choice of two different methods for temperature control (an external circulating waterbath for heating and cooling, or a built-in electric heater).

1. Computer-operated control with Touch Screen Interface

2. Pressure Range: ambient - 45,000psi (310 MPa)

3. Choice of temperature control - heating/cooling or heating only

4. Max Temperature: 95˚C

5. Min Temperature: 4˚C with circulating waterbath, ambient with electric heating

6. Chamber capacity: up to 16 samples in MicroTubes

7. Easily accessible USB port on front panel

8. Air pressure: >0.8 Mpa. Input air pressure can be provided by an air compressor, house air or nitrogen lines, air or nitrogen tanks.

9. Pressure medium: Distilled Water

Workflow

Minute sample preparation protocol

PCT Authorized Distributors :

1.Zhengzhou Jiemeng Instrument & Equipment Co., Ltd.

2.TOCH Development Shenzhen Co., Ltd.

3.Shanghai Dynamax Biotech Co., Ltd.

4.Fuzhou Zkcreate Biotech Co., Ltd.

5.Shenyang Ruibang Biotechnology Co., Ltd.

6.Beijing Qiangxin Biorepublic Co., Ltd.

7.Hangzhou baiaofa Biotechnology Co., Ltd.

References:

1.Szabo et al. Rapid release of N-linked glycans from glycoproteins by pressure-cycling technology. Anal Chem. 2010.82(6):2588-2593

https://pubs.acs.org/doi/10.1021/ac100098e

2.Washburn et al. Controlled tetra-Fc sialylation of IVIg results in a drug candidate with consistent enhanced anti-inflammatory activity. Proceedings of the National Academy of Sciences. 2015.112(11):1297-1306

https://www.pnas.org/content/112/11/E1297

3.Guo et al. Rapid mass spectrometric conversion of tissue biopsy samples into permanent quantitative digital proteome maps. Nature Medicine.2015.21:407-413

https://www.nature.com/articles/nm.3807

4.Jelcic et al. Memory B Cells Activate Brain-Homing, Autoreactive CD4(+) T Cells in Multiple Sclerosis. Cell. 2018.175(1):85-100

https://www.cell.com/cell/fulltext/S0092-8674(18)31027-4

5.Xuan, et al. Standardization and harmonization of distributed multi-center proteotype analysis supporting precision medicine studies. Nat Commun. 2020.11(1):1-12

https://www.nature.com/articles/s41467-020-18904-9

6.Gao et al. Accelerated Lysis and Proteolytic Digestion of Biopsy-Level Fresh-Frozen and FFPE Tissue Samples Using Pressure Cycling Technology. J Proteome Res. 2020.19(5):1982-1990

https://pubs.acs.org/doi/10.1021/acs.jproteome.9b00790

7.Poulos et al. Strategies to enable large-scale proteomics for reproducible research. Nat Commun. 2020.11(1):1-13

https://www.nature.com/articles/s41467-020-17641-3

8.Nie et al.Multi-organ proteomic landscape of COVID-19 autopsies. Cell. 2021.184(3):775-791

https://www.cell.com/cell/fulltext/S0092-8674(21)00004-0

9.Zhu et al. SnapShot: Clinical proteomics. Cell.2021.184(18):4840

https://www.cell.com/cell/fulltext/S0092-8674(21)00985-5

10.Tanet et al. DNA transposons mediate duplications via transposition-independent and -dependent mechanisms in metazoans. Nature Communication. 2021.12(1):4280

https://www.nature.com/articles/s41467-021-24585-9

PCT (Pressure Cycling Technology), a rapid sample pre-treatment method for micro-scale number of samples, is of high robustness. It is a patented technology platform based on repeated cycles of hydrostatic pressure between ambient (14.69 PSI) and ultra-high (45,000 PSI) levels. These rapid changes in pressure are used to control biomolecular interactions for applications such as accelerated proteolytic digestion, and improved extraction of cellular components, including proteins and lipids.

The PCT-HD Sample Preparation System is a proven PCT-based workflow for processing small tissue samples for proteomic and other applications. PCT-HD combines efficient, hands-off tissue homogenization and protein extraction with pressure-enhanced digestion, for rapid, efficient and reproducible generation of peptides for MS analysis. This unique workflow combines two of PBI’s innovative sample preparation tools: the Barocycler 2320EXT and our patented PCT MicroPestles. PCT-HD and the Barocycler 2320EXT can provide unprecedented speed and reproducibility for biomarker discovery, yielding peptides ready for clean-up and MS within 2-4 hours from the start of tissue processing.

The Barocycler 2320EXT is a compact, bench top instrument that can process up to 16 samples simultaneously using PCT MicroTubes. Features that come standard with the 2320EXT include: data input and output options to accommodate validation and quality control; computer-operated control with touch screen programming and automatic data logging; and the ability to control multiple pressure cycling parameters, such as the rate of pressure increase/decrease, both the high and low pressure levels, and the shape of the pressure profile (e.g., sine wave, square wave, and others). In addition, the 2320EXT is available with a choice of two different methods for temperature control (an external circulating waterbath for heating and cooling, or a built-in electric heater).

1. Computer-operated control with Touch Screen Interface

2. Pressure Range: ambient - 45,000psi (310 MPa)

3. Choice of temperature control - heating/cooling or heating only

4. Max Temperature: 95˚C

5. Min Temperature: 4˚C with circulating waterbath, ambient with electric heating

6. Chamber capacity: up to 16 samples in MicroTubes

7. Easily accessible USB port on front panel

8. Air pressure: >0.8 Mpa. Input air pressure can be provided by an air compressor, house air or nitrogen lines, air or nitrogen tanks.

9. Pressure medium: Distilled Water

Workflow

Minute sample preparation protocol

References:

1.Szabo et al. Rapid release of N-linked glycans from glycoproteins by pressure-cycling technology. Anal Chem. 2010.82(6): 2588-2593

https://pubs.acs.org/doi/10.1021/ac100098e

2.Washburn et al. Controlled tetra-Fc sialylation of IVIg results in a drug candidate with consistent enhanced anti-inflammatory activity. Proceedings of the National Academy of Sciences. 2015.112(11): 1297-1306

https://www.pnas.org/content/112/11/E1297

3.Guo et al. Rapid mass spectrometric conversion of tissue biopsy samples into permanent quantitative digital proteome maps. Nature Medicine.2015.21:407-413

https://www.nature.com/articles/nm.3807

4.Jelcic et al. Memory B Cells Activate Brain-Homing, Autoreactive CD4(+) T Cells in Multiple Sclerosis. Cell. 2018.175(1):85-100

https://www.cell.com/cell/fulltext/S0092-8674(18)31027-4

5.Xuan, et al. Standardization and harmonization of distributed multi-center proteotype analysis supporting precision medicine studies. Nat Commun. 2020.11(1):1-12

https://www.nature.com/articles/s41467-020-18904-9

6.Gao et al. Accelerated Lysis and Proteolytic Digestion of Biopsy-Level Fresh-Frozen and FFPE Tissue Samples Using Pressure Cycling Technology. J Proteome Res. 2020.19(5):1982-1990

https://pubs.acs.org/doi/10.1021/acs.jproteome.9b00790

7.Poulos et al. Strategies to enable large-scale proteomics for reproducible research. Nat Commun. 2020.11(1):1-13

https://www.nature.com/articles/s41467-020-17641-3

8.Nie et al.Multi-organ proteomic landscape of COVID-19 autopsies. Cell. 2021.184(3):775-791

https://www.cell.com/cell/fulltext/S0092-8674(21)00004-0

9.Zhu et al. SnapShot: Clinical proteomics. Cell.2021.184(18):4840

https://www.cell.com/cell/fulltext/S0092-8674(21)00985-5

10.Tanet et al. DNA transposons mediate duplications via transposition-independent and -dependent mechanisms in metazoans. Nature Communication. 2021.12(1):4280

https://www.nature.com/articles/s41467-021-24585-9

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