Precise characterization of a monolayer of two different biomolecules in a gradient pattern on a glass surface puts high demand on the method used. Some techniques can detect protein monolayers but not on a glass surface. Others can distinguish between different proteins but not identify a gradient pattern. Here, we used ToF-SIMS to validate the complete surface composition, checking all the necessary boxes. As these types of surfaces can dictate sensitive cell behaviors, the precision on a nanolevel is crucial, and to visualize and determine the molecular distribution become essential. The dual monolayer consisted of laminin 521 and one of three other biomolecules of different sizes, epidermal growth factor, growth differentiation factor 5, or bovine serum albumin, creating opposing gradient patterns. The resulting ToF-SIMS imaging and line scan data provided detailed information on the distribution of the adsorbed proteins.

1.
E. H.
Tronic
,
Surface Analysis of Adsorbed Proteins: A Multi-Technique Approach to Characterize Surface Structure
(
University of Washington
, Seattle, WA,
2012
).
2.
C.
Gruian
,
E.
Vanea
,
S.
Simon
, and
V.
Simon
,
Biochim. Biophys. Acta Proteins Proteomics
1824
,
873
(
2012
).
3.
F.
Höök
,
B.
Kasemo
,
T.
Nylander
,
C.
Fant
,
K.
Sott
, and
H.
Elwing
,
Anal. Chem.
73
,
5796
(
2001
).
4.
R.
Konradi
,
M.
Textor
, and
E.
Reimhult
,
Biosensors
2
,
341
(
2012
).
5.
A.
Lundgren
,
M.
Hulander
,
J.
Brorsson
,
M.
Hermansson
,
H.
Elwing
,
O.
Andersson
,
B.
Liedberg
, and
M.
Berglin
,
Part. Part. Syst. Charact.
31
,
209
(
2014
).
6.
E.
Dulkeith
 et al,
Phys. Rev. Lett.
89
,
203002
(
2002
).
7.
J.
Borglin
,
S.
Guldbrand
,
H.
Evenbratt
,
V.
Kirejev
,
H.
Grönbeck
, and
M. B.
Ericson
,
Appl. Phys. Lett.
107
,
234101
(
2015
).
8.
Q. P.
Vanbellingen
,
N.
Elie
,
M. J.
Eller
,
S.
Della-Negra
,
D.
Touboul
, and
A.
Brunelle
,
Rapid Commun. Mass Spectrom.
29
,
1187
(
2015
).
9.
P.
Kingshott
,
G.
Andersson
,
S. L.
McArthur
, and
H. J.
Griesser
,
Curr. Opin. Chem. Biol.
15
,
667
(
2011
).
10.
A. M.
Belu
,
D. J.
Graham
, and
D. G.
Castner
,
Biomaterials
24
,
3635
(
2003
).
11.
P.
Malmberg
,
T.
Guttenberg
,
M. B.
Ericson
, and
L.
Hagvall
,
Contact Dermatitis
78
,
109
(
2018
).
12.
P.
Malmberg
,
E.
Jennische
,
D.
Nilsson
, and
H.
Nygren
,
Anal. Bioanal. Chem.
399
,
2711
(
2011
).
13.
B.
Li
,
Y.
Ma
,
S.
Wang
, and
P. M.
Moran
,
Biomaterials
26
,
1487
(
2005
).
14.
R.
Michel
and
D. G.
Castner
,
Surf. Interface Anal.
38
,
1386
(
2006
).
15.
M.
Hulander
,
A.
Lundgren
,
M.
Berglin
,
M.
Ohrlander
,
J.
Lausmaa
, and
H.
Elwing
,
Int. J. Nanomed.
6
,
2653
(
2011
).
16.
A. M.
Lipski
,
C. J.
Pino
,
F. R.
Haselton
,
I.-W.
Chen
, and
V. P.
Shastri
,
Biomaterials
29
,
3836
(
2008
).
17.
K.
Anselme
,
L.
Ploux
, and
A.
Ponche
,
J. Adhes. Sci. Technol.
24
,
831
(
2010
).
18.
A.
Lundgren
,
Y.
Hed
,
K.
Öberg
,
A.
Sellborn
,
H.
Fink
,
P.
Löwenhielm
,
J.
Kelly
,
M.
Malkoch
, and
M.
Berglin
,
Angew. Chemie Int. Ed.
50
,
3450
(
2011
).
19.
L.
Andreasson
,
H.
Evenbratt
, and
S.
Simonsson
,
Heliyon
6
,
e04133
(
2020
).
20.
L.
Andreasson
,
H.
Evenbratt
,
R.
Mobini
, and
S.
Simonsson
,
J. Biotechnol.
(published online, 2020).
21.
M.
Arnold
 et al,
Nano Lett.
8
,
2063
(
2008
).
22.
R. J.
McMurray
 et al,
Nat. Mater.
10
,
637
(
2011
).
23.
M. J.
Dalby
,
N.
Gadegaard
,
R.
Tare
,
A.
Andar
,
M. O.
Riehle
,
P.
Herzyk
,
C. D. W.
Wilkinson
, and
R. O. C.
Oreffo
,
Nat. Mater.
6
,
997
(
2007
).
24.
G.
Maheshwari
,
G.
Brown
,
D. A.
Lauffenburger
,
A.
Wells
, and
L. G.
Griffith
,
J. Cell Sci.
113
,
1677 LP
(
2000
).
25.
M. P.
Lutolf
and
J. A.
Hubbell
,
Nat. Biotechnol.
23
,
47
(
2005
).
26.
J. B.
Gurdon
and
P.-Y.
Bourillot
,
Nature
413
,
797
(
2001
).
27.
28.
B. J.
Tyler
,
C.
Bruening
,
S.
Rangaranjan
, and
H. F.
Arlinghaus
,
Biointerphases
6
,
135
(
2011
).
29.
R. N.
Foster
,
E. T.
Harrison
, and
D. G.
Castner
,
Langmuir
32
,
3207
(
2016
).
30.
M. S.
Kim
,
G.
Khang
, and
H. B.
Lee
,
Prog. Polym. Sci.
33
,
138
(
2008
).
31.
See supplementary material at https://doi.org/10.1116/6.0000621 for further information on protein structure, spectra, etc.

Supplementary Material

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