News & Featured Publications

2023 New Year’s Promotion

January 3, 2023

Happy New Years 2023

New Year’s promotion: 10% Off Protein Identification (Offer Ends 02/03/2023)

Happy 2023! We would like to show our appreciation for choosing us to serve your proteomics needs. From January 3 to February 3, we are offering 10% off protein identification of 2D gel spots or 1D gel bands.

Take advantage of this limited-time discount to kick start your research for the new year. This offer applies to all our loyal existing customers as well as new customers.

*Terms: Only applies for Protein ID by standard MALDI-TOF/TOF, not LC-MS/MS. Cost of running 2D DIGE gel not covered. Does not apply to previous orders. Applied Biomics’ standard service terms and conditions apply.

Identify Critical Phospho-Sites using our Phospho-Site identification service

November 17, 2022

A recent study in eLife used Applied Biomics’ Phospho-Site identification service to identify critical phospho-sites of A-kinase anchoring protein 12 (AKAP12) involved in liver fibrosis.

Activation of hepatic stellate cells (HSCs) causes liver fibrosis by releasing extracellular matrix. Although previous work has shown an association between the phosphorylation of AKAP12 and HSC activation, the specific phosphorylation sites remain largely unknown.

Identification of phosphorylation-site involves enriching phosphorylated peptides followed by nanoLCMSMS. The presence of phospho-sites is confirmed by the presence of a MSMS peak showing the loss of phosphate (circled red).

From the MSMS spectra, multiple AKAP12 phosphorylation sites in activated HSC samples were identified and listed in the following table:

Interestingly, if these phosphor-sites were edited by CRISPR, the ability of AKAP12 to inhibit HSC activation is restored. The authors showed that AKAP12 phospho-editing dramatically inhibits fibrosis, ER stress response, HSC inflammatory signaling, and liver injury in mice.

Comments: This publication has clearly shown the significant role that phosphorylation could play. However, it is very challenging to study phosphorylation and other post translational modifications (PTMs) due to the very low level of these PTMs in vivo. In this project, we enriched the phosphorylated peptides and used the mass spectrometry settings for PTM detection, which enhanced detection sensitivity significantly.

If you are interested in a similar study, feel free to contact our support team with a brief description of your project. Our scientists will reach out to you shortly.

Track HCP profile during bioprocess

October 18, 2022

Applied Biomics offers comprehensive HCP analysis to detect and quantify HCP profiles during the purification process:

1. Qualitative Analysis: First, customers can visualize HCP compositions, modifications and fragmentations in a high resolution, large 2D image. This cannot be achieved by ELISA, LC-MSMS or other platforms.

2. Quantitative Analysis: Next, the HCPs can be analyzed quantitatively as following:

  • Number of total detected spots in each sample
  • Number of unique and overlapping spots between samples
  • HCP quantitation: quantify individual HCP or total HCP in each sample. 
  • Spot map: Below is an example with 2-fold cutoff comparing CHO_S1 vs. CHO_S2. Red, green and yellow indicate spots with increased>2-fold, decreased>2-fold and within 2-fold, respectively 

Please view here for a detailed example

3. Wide range of applications:

We have further optimized our proprietary sample preparation protocols for all major organisms used for generating recombinant therapeutic proteins. This assay can achieve 0.1 ng/spot sensitivity, equivalent to 1 PPM with 0.1 mg total protein loading. Quantitation of fluorescent signal allows accurate quantitation of the HCP and DS.

4. Cost-Effective

Lastly, each assay simultaneously tests and compares up to 3 samples, offering great saving of cost.

Featured Publication: Kinase Substrate Mapping Using Phospho-Proteomics Platform

September 19, 2022

A recent publication in Microbiology Spectrum used our PhosphoProteomics platform to identify important kinase substrates forProtein Kinase K (PknK) in Mycobacterium tuberculosis.

Pknk plays a key role in regulating the growth of M. tuberculosis. The PknK knock out strain showed increased survival under metabolic stress such as carbon and nitrogen starvation signals, compared to WT strains.

To identify Pknk substrates, authors compared the global phosphoproteomic profiles of WT and phosphorylation-defective PknK strains. Both the phosphorylation and protein level of each protein was assessed. Among thousands of proteins, only 22 spots showed positive phosphorylation level in the WT but not the Pknk-defective strain, as well as protein level change.

Further mass spectrometry of the 18 spots identified Pknk substrates such as Rho transcription terminator and several critical enzymes involved in metabolism and signal transduction.

Together with other findings, this study further established the Pknk signal transduction network and its central role for mycobacterial survival.

Comments: It is challenging to study protein phosphorylation due to the low phosphor-level and lack of effective separation between phosphor-protein and native protein. Our PhosphoProteomics platform addresses both these issues through fast screening and quantification of phospho-proteins with high sensitivity and resolution. The authors in this publication were able to quickly narrow down their targets to 22 protein spots among thousands of proteins. More importantly, proteins identified using our platform were validated by transcriptome and in vivo analysis.

Featured Service: Choosing the right sample type for HCP Antibody Coverage

August 17, 2022

It is critical to detect and measure HCPs in therapeutic products. However, a common drawback in many assays is the inability to detect protein modifications and degradations.

Our HCP Antibody Coverage platform can detect HCP composition, protein modifications, and degradations with high sensitivity and consistency – making it advantageous to ELISA, AAE and standard 2D Western blot. These features can greatly improve the purification steps during manufacturing and purity analysis of the final drug substance (DS).

While high quality HCP antibody is critical for optimal HCP coverage, it is equally important to determine the correct sample type: host cell culture supernatant, cell pellets, or others.

Here, we tested HCP antibody coverage of the same antibody against CHO cell supernatant (CHO-S) and CHO cell pellets (CHO-P). A striking difference is observed: 97.1% for CHO-S and 70.0% for CHO-P.

If you are interested, feel free to contact our support team with a brief description of your project. Our scientists will reach out to you shortly.

Featured Publication: Tissue Barriers

July 20, 2022

Fig 1b: 2-D gel separation of proteins present in non-magnetic and magnetic capture membrane fractions with location of spot 16 identified by mass spectrometry as GRP75.

A recent publication in Tissue Barriers used our 2D DIGE / Mass Spec platform to identify key proteins in toxin-induced cell death.

Cholix is an exotoxin which induces cell death after crossing the epithelium. After comparing the protein profile of Cholix magnetic capture samples to the whole cell membrane (control) by 2D DIGE, 45 proteins showed significant differences. All 45 proteins were identified by mass spectrometry. One of the most over-expressed proteins in Cholix magnetic capture samples was GRP75 (spot 16).

GRP75, a heat shock protein that plays an important role in vesicles, was found to interact with Cholix during its crossing of the epithelium. Following endocytosis, GRP75 limits the delivery of Cholix to lysosomes.

More importantly, the binding of GRP75 to Cholix was further confirmed by 2 other platforms: IP and ELISA.

Comments on the data: With the fast development of proteomics technologies, researchers are often facing the challenge of identifying ‘real’ biomarkers that can be validated by other platforms. The findings in this paper proved again that 2D DIGE is a reliable platform to analyze complex proteomes and identify biomarkers.

If you are interested, feel free to contact our support team with a brief description of your project. Our scientists will reach out to you shortly.

Featured Service: Improved HCP antibody coverage protocol

June 15, 2022

Brown: 3015 spots detected by anti-CHO HCP antibody
Blue: 83 spots not detected by antibody
Anti-CHO HCP antibody coverage: 97.3%

One main challenge in performing HCP antibody coverage by 2D Western blot is the unspecific antibody binding, which causes high background and inaccuracy in quantitation. To address this issue, Applied Biomics has performed extensive internal studies testing different conditions.

With our recent improvements to the protocol, we were able to achieve:

  1. Significantly lower background
  2. Overall higher HCP antibody coverage

Featured Publication: bioRxiv

May 18, 2022

Study protein distribution within cells using our mass spectrometry service. A recent bioRxiv paper used Mass Spectrometry to identify proteins in various sections of a giant cell. Findings revealed that 30% of the Stentor proteome has polarized localization.

Featured Service: PTMs by Mass Spectrometry

April 19, 2022

Interested in PTM sites?

Our mass spectrometry service can identify PTM sites such as phosphorylation, acetylation, methylation (and more) with high sensitivity and accuracy. The service also includes the confirmation by MS/MS spectrum showing the presence of signature ions (if applicable).

Featured Publication: Int J Biol Sci

March 16, 2022

Identify important breast cancer biomarkers using our 2D DIGE service

An Int J Biol Sci study demonstrated the up-regulation of G6PD in doxorubicin-resistant breast cancer cells. Findings suggest that G6PD inhibition can be a promising strategy against this cancer.

Featured Publication: Cell

February 16, 2022

Quantitative neuronal proteome analysis using our iTRAQ services:

A study published in Cell quantifies proteome changes between control and chaperone-mediated autophagy (CMA) deficient mice using iTRAQ. The findings showed that functional CMA is essential for neuronal proteostasis to prevent neuro-degeneration.

News Release: Applied Biomics Offers Qualitative and Quantitative Host Cell Protein (HCP) Profiling Analysis

February 19, 2020

From Business Wire:

Applied Biomics, Inc., a leading Proteomics service provider, adds a HCP profiling analysis using their well established high resolution, large format 2-Dimensional Differential In-Gel Electrophoresis (2D DIGE) platform. This assay can be used to monitor HCP contamination during the purification process thus helps to improve purification strategy, specifically:

1) Visualize and overlay the HCP images between different samples
2) Quantitate HCP content by spot counting and PPM for each sample
3) Quantitate the purity of product or drug substance (DS) by spot counting and PPM
4) Quantitatively compare HCP content by fold-change and overall similarity between samples