1000 proteins quantified in e. coli mock lysate - HCP

Is there a simple way to perform label-free quantification of Host Cell Proteins?

Question: I find it difficult to get an accurate label-free quantification of Host Cell Proteins with LC-MS. It also seems that...

Jun 25. 2018 | Ejvind Mørtz | 0

What are “Jackpot HCPs” and why do I need a “lucky goat” to get a precise result in my HCP ELISA?

Question: I tested two different HCP ELISA kits on my biopharmaceutical. Why do they give different results and which one is...

May 28. 2018 | Anette Draborg | 0

Quickly compare HCP in different process steps, without setting up an ELISA

Question: I have spent days trying to set up a useful ELISA assay for detecting Host Cell Proteins (HCPs) in my process samples. I'm...

Apr 10. 2018 | Ejvind Mørtz | 0
HPLC analysis of process related impurities by reverse phase mass spectrometry ( LC-MS analysis )

How to identify peaks observed by UV-HPLC in stability studies

Question: We have developed a Reversed-Phase (RP) UV- HPLC chromatography method. It is used for stability studies of our new pharmaceutical protein. How do we...

Jan 24. 2018 | Thomas Kofoed | 0
What is SWATH?

What is SWATH and why is it an advantage for Host Cell Protein analysis?

Host Cell Proteins (HCPs) in protein biopharmaceuticals can be analyzed by mass spectrometry - for both identification and quantification of...

Jun 22. 2017 | Thomas Kofoed | 0

4 reasons why Amino-terminal sequencing of mAbs should be done by Edman degradation

Question: How do I obtain the N-terminal sequences of the heavy chain and light chain of my monoclonal antibody?   Answer: mAb N-terminal sequence...

Apr 26. 2017 | Ejvind Mørtz | 0
Protein calculator

GPMAW lite – Protein Molecular Weight Calculator (and much more)

Simple bioinformatics calculations on specific protein sequences When working with proteins you often need to make simple bioinformatics calculations on a specific...

Apr 10. 2017 | Thomas Kofoed | 0
Antibody characterization

How to: Make sure that Ile/Leu are correctly assigned by de-novo sequencing of your antibody

Question: I was told that de-novo sequencing by mass spectrometry is not able to distinguish between Ile and Leu residues in my...

Mar 22. 2017 | Fen Yang Reske-Nielsen | 0
Application note - mAb

What purity is required for de-novo sequencing of my antibody?

Question: My monoclonal antibody is not 100% pure, what is the purity required for de-novo sequencing?   Answer: We typically observe two types of protein impurities in...

Mar 20. 2017 | Fen Yang Reske-Nielsen | 0
100% sequence coverage map of antibody

Antibody de-novo Sequencing by Mass Spectrometry: How to get 100% sequence coverage

Question: Is it possible to perform a de-novo sequencing of my purified monoclonal antibody without the hybridoma cells?   Answer: Yes! – Your purified antibody can...

Mar 17. 2017 | Fen Yang Reske-Nielsen | 0

Reasons why you need accurate concentration determination of your protein standards

How do I quantify my protein standard? Have you ever questioned the quantitative data you get from Bradford, Lowry or BCA protein quantification...

Jan 18. 2017 | Thomas Kofoed | 0
Extinction coefficient protein concentration

How to determine the Molar Extinction Coefficient of your protein (in 3 small steps)

Question: I have a purified protein that I would like to quantify accurately in my lab using 280 nm UV measurement...

Nov 15. 2016 | Thanh Ha Nguyen | 0
Analysis method for Mw HCPs (small HCPs) below 20 kDa

Identification of small host cell proteins (low Mw HCPs)

Question: Could you suggest an analysis method to detect low Mw HCPs (small HCPs) below 20 kDa? I work on a drug...

Oct 11. 2016 | Ejvind Mørtz | 0
Amino Acid analysis

4 things to remember about Amino Acid Analysis of proteins and peptides

Amino acid analysis is a method to determine the absolute amounts of individual amino acids in a sample. The method...

Sep 23. 2016 | Ejvind Mørtz | 1
2D PAGE preparation for MALDI ISD

1D SDS PAGE versus 2D PAGE. I’m considering 2D electrophoresis, any recommendations?

Question: I’m interested in isolating and identifying cell surface receptor proteins. They typically have high Mw, are glycosylated, and also transmembrane...

Sep 18. 2016 | Ejvind Mørtz | 0
Protein scientist at Alphalyse

3 steps to identify and quantify HCP in a drug substance homolog from a host cell

Question: My HCP antibodies show a reaction towards a drug substance band in my Western blot. Is it possible that it is...

Sep 5. 2016 | Rikke Raaen Lund | 0

How should I prepare my samples for 2D electrophoresis, and how much protein should I load to get protein identification by mass spec?

How should I prepare my samples for 2D electrophoresis, and how much protein should I load to get protein ID...

Jun 8. 2016 | Ejvind Mørtz | 0
N- and C-terminal sequencing from PVDF membrane

How to increase protein concentration before loading onto 1D SDS PAGE

Question: I have another question:  Is it ok if I precipitate my samples with TCA prior loading to the SDS-Gel? If not, how...

Jun 7. 2016 | Ejvind Mørtz | 0
Mass spectrometry analysis of protein

Common protein contaminants observed in mass spectrometric protein ID

Questions: Is the keratin identified by the protein ID service in my 1D gel band a contamination? Or is it...

Jun 3. 2016 | Ejvind Mørtz | 0

Effective 1D/2D PVDF staining protocols (for N-terminal sequencing)

For N-terminal Edman sequencing SDS PAGE separated proteins can be electroblotted onto PVDF membranes. After PVDF staining You may cut...

May 24. 2016 | Ejvind Mørtz | 0
Eppendorf tubes with protein

Mass spectrometric identification of proteins detected in Western blots

Question: Dear Sir, We have an unknown protein recognized in Western blot by autoantibodies that we need to identify. Is it...

May 2. 2016 | Ejvind Mørtz | 0
Preparation of sample for MALDI

What buffer to use for MALDI mass spectrometry analysis of a protein?

Question: What buffer should I use for MALDI Mass Spectrometry analysis of my protein sample? Answer: In general, MALDI-MS is more tolerant...

Apr 28. 2016 | Ejvind Mørtz | 1

Useful Protein Molecular Weight Calculator kDa

Protein molecular weight calculator kDa The rule of thumb for conversion of microgram amounts into picomole amounts at different protein molecular...

Nov 10. 2015 | Ejvind Mørtz | 0

What does the Mascot Score mean?

Question: I just got my protein identification report. What does the Mascot Score mean? Answer: The Protein Identification report contains a list of proteins that have...

Oct 9. 2015 | Ejvind Mørtz | 0

Why is my protein’s N-terminal blocked (and what can I do about it)?

N-terminal sequencing is a requirement according to the ICH Q6B Guideline. This includes structural characterization of recombinant proteins for clinical...

Aug 31. 2015 | Ejvind Mørtz | 2

Identification of proteolytic degradation of recombinant protein

Question: I purified a recombinant protein and analyzed it by 1D SDS PAGE. In addition to the intact protein (52...

May 9. 2015 | Ejvind Mørtz | 2

Guide to identification of novel proteins not present in public databases

Question: I recently used Alphalyse's protein identification service to analyze my protein. The sample gave me very good MS data. However it...

Dec 2. 2014 | Ejvind Mørtz | 0

Preparing samples for N- and C-terminal sequencing by MALDI ISD?

Question: I am interested in your N and C-terminal sequencing using MALDI ISD. How do I prepare the sample for...

Feb 22. 2014 | Ejvind Mørtz | 0

Assessing the stability of a protein drug substance and formulated drug product

After protein purification, an important issue is the stability of a protein drug substance and formulated drug product. Stability assays...

Oct 21. 2013 | Ejvind Mørtz | 1

How to optimize electroblotting conditions onto PVDF membrane for N-terminal Edman sequencing

How do I optimize electroblotting conditions onto PVDF membrane for N-terminal Edman sequencing? Our 12 kDa protein separates well on...

Oct 13. 2013 | Ejvind Mørtz | 0
N- and C-terminal Sequencing

3 Techniques for N- and C-terminal sequencing of proteins

Where does the amino acid sequence start and where does it end? The terminal amino acid sequences are important for the...

Sep 16. 2013 | Ejvind Mørtz | 2

N- and C-terminal protein sequencing using MALDI ISD

Structural characterization of purified natural and recombinant proteins traditionally includes N-terminal Edman sequencing to confirm 5-15 amino acid residues from...

Aug 18. 2013 | Ejvind Mørtz | 0
Order analysis online with quick response

Which databases and organisms are used for mass spec protein ID service?

Question: I would like to identify proteins from an organism where the genome has just been sequenced. How many proteins are...

Nov 16. 2009 | Ejvind Mørtz | 0

Mass Spectrometry for Host Cell Protein monitoring and control in pharmaceutical process development

Mass spectrometry based Host Cell Protein identification and quantification Here the Mass Spec Host Cell Protein (HCP) assay is applied for...

Oct 2. 2009 | Ejvind Mørtz | 2

Protocol for silver staining of gels optimized for mass spectrometry and protein identification

Problems arising from using commercial staining kits: Scientists use silver staining for sensitive detection of proteins separated by 1D and 2D...

Aug 6. 2009 | Ejvind Mørtz | 1
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