Oxidation of Methionine

D Y M P K

O

N-terminal Glutamine to pyroGlu

Q V Q L V

O

O

R

H2N NH

R + H2O O

Conjugation sites

D A C P K

N-linked

O-linked

N Y S T K

S D Y C K

Glycosylation

A C T Y K

A C D Y K

Sequence Variance

D C D Q K

A Y C S K

S-S

Disulfide Bridge location

P G K

P G

C-terminal Lysine clipping

O

β

α

O

NH

NHOH

isoAspD or

P E N N Y K

Deamidation of Asparagine

Points of interest on a peptide map

What do we look for when mapping biopharmaceuticals?

RT

A

C

D

Y

M

D

K

A

C

D

Y

K

Y

M

D

K

D

P

C

Y

K

C

D

K

Peptide Separation

m/z

A

C

D

Y

M

D

K

A

C

D

Y

K

Y

M

D

K

D

P

C

Y

K

C

D

K

Peptide Detection

A C D Y M D K

A C D Y K

Y M D K D P C Y K

C D K

N-terminus –

– C-terminus

Data Interpretation

Protein Digestion A C D Y K D P C Y K

Mapping a biopharmaceutical route

What are the steps involved in peptide mapping?

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Learn more at: thermofisher.com/PeptideMaps

• Complete and reproducible protein digestion

• High resolution reproducible peptide separations

• HRAM MS and MS/MS

• Powerful yet user-friendly software

Choosing the correct tools for your journey

What are the key tools to confidently map a protein?

• Peptide mapping has the selectivity to simultaneously monitor many attributes or points of interest.

• This is important for maintaining the safety and efficacy of protein therapeutics