Table 3 Classification of mtDod-PAS-Pep derived ABs. ABs were classified as followed: “Class 1” ABs recognize the proteins of interest provided as recombinantly purified protein (produced in E. coli). “Class 2” ABs do not recognize recombinant protein of interest, but protein of expected apparent molecular weight in HEK293T human cell lysates. “Class 3” ABs only recognize mtDod-PAS constructs (like the respective mtDod-PAS-Pep or mtDod-PAS-Met). Western blots of ABs rated as “class 1” or “class 2” are shown in Fig. 8. The AB derived from mtDod-PAS-Pep6 and mtDod-PAS-Pep8 only recognized mtDod-PAS-Pep constructs and were considered “class 3” (Supplementary Fig. S11).

From: Dodecin as carrier protein for immunizations and bioengineering applications

ABs derived from mtDod construct

Binding region of the ABs

AB class

mtDod-PAS-Pep1

HSP-70

C-terminus

Class 1

mtDod-PAS-Pep2

HSP-90

C-terminus

Class 1

mtDod-PAS-Pep3

HSP-110

C-terminus

Class 1

mtDod-PAS-Pep4

HSP-A4

C-terminus

Class 1

mtDod-PAS-Pep5

H3

C-terminus

Class 2*

mtDod-PAS-Pep6

H4

C-terminus

Class 3

mtDod-PAS-Pep8

HB-EGF**

Pos. 20–34

Class 3

mtDod-PAS-Pep9

CHIP

N-terminus

Class 1

mtDod-PAS-Pep10

CHIP

Broken helix

Pos. 151–161

Class 2***

mtDod-PAS-Pep11

CHIP

Tip of helix

Pos. 183–192

Class 1

  1. *mtDod-PAS-Pep5 derived ABs showed only a very weak signal for 1 µg and 500 ng of recombinant protein with no intensity difference (see Supplementary Fig. S11). Thus, the AB preparation was regarded as “class 2”.
  2. **Proheparin-binding EGF-like growth factor of Chlorocebus aethiops (green monkey).
  3. ***mtDod-PAS-Pep10 derived ABs didn’t recognize purified CHIP but seem to recognize a protein in CHIP-overexpressing cells; no detection range was determined.
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