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Table 1 Expression of PfEMP1 domains in P. pastoris

From: Optimizing expression of the pregnancy malaria vaccine candidate, VAR2CSA in Pichia pastoris

Gene Domain Parasite strain construct MW1 Protein Secreted IE surface reactivity2
var2CSA DBL1 IT/FCR3 3 H58 – S383 38.9 ++ yes
var2CSA DBL1 3D7 H58 – I433 45.2 ++ yes
var2CSA DBL1 7G8 H57-I437 46.3 ++ yes
var2CSA DBL1 Dd2 H57-I433 45.6 ++ yes
var2CSA ID1-DBL2 IT/FCR3 V490 – D883 44.5 + nt
var2CSA DBL2 IT/FCR3 3 K543 – K838 36.1 - nt
var2CSA DBL2 IT/FCR3 D517 – D883 41.6 + nt
var2CSA DBL2 IT/FCR3 D517 – L1024 58.4 - nt
var2CSA DBL2 IT/FCR3 G550 – L1024 54.6 - nt
var2CSA DBL2 IT/FCR3 D545 – L1024 55.2 - nt
var2CSA DBL2-ID2 IT/FCR3 D517 – G1229 81.6 - nt
var2CSA ID1-DBL2-ID2 IT/FCR3 V490 – G1229 84.4 - nt
var2CSA ID2 IT/FCR3 V878 – K1201 38.4 - nt
var2CSA ID2 IT/FCR3 S843 – G1229 45.2 - nt
var2CSA ID2 IT/FCR3 L1024 – G 1229 24.9 - nt
var2CSA ID2-DBL3 IT/FCR3 N 859 – C 1576 83.0 - nt
var2CSA DBL3 IT/FCR3 3 L1221 – E1541 37.3 negligible yes
var2CSA DBL3 IT/FCR3 C1202 – C1576 43.8 + yes
var2CSA DBL3 3D7 C1224 – Y1510 33.2 - nt
var2CSA DBL3 Dd2 C1219 – Y1529 44.2 - nt
var2CSA DBL3 Pc49 C1227 – Y1540 43.8 - nt
var2CSA DBL3 7G8 C1199 – E1587 45.0 ++ nt
var2CSA DBL4 IT/FCR3 3 S1594 – V1888 34.7 ++ no4
var2CSA DBL4-DBL5 IT/FCR3 G1588 – E2288 85.4 - nt
var2CSA DBL4-DBL5 IT/FCR3 S1594 – E2288 82.5 - nt
var2CSA DBL4-ID4-DBL5 IT/FCR3 S1594 – R2271 66.8 - nt
var2CSA DBL4-DBL5 3D7 S1576 – T2291 85.6 - nt
var2CSA DBL4-DBL5 3D7 G1550 – T2291 88.3 - nt
var2CSA DBL4-DBL5 7G8 S1604 – T2318 89.2 - nt
var2CSA DBL4-DBL5 7G8 G1578 – T2318 88.3 + nt
var2CSA DBL5 IT/FCR3 3 L2003 – L2270 31.7 - nt
var2CSA DBL5 IT/FCR3 N1893 – N2290 46.6 - nt
var2CSA DBL5 IT/FCR3 K1984 – E2288 36.6 + yes
var2CSA DBL5 IT/FCR3 E1971 – N2290 37.8 - nt
var2CSA DBL5 7G8 Q2000 – T2318 38.8 ++ yes
var2CSA DBL5 3D7 L1997 – A2269 32.2 - nt
var2CSA DBL5 3D7 K1978 – T2291 37.9 - nt
var2CSA DBL5 3D7 L1997 – T2291 35.8 - nt
var2CSA DBL5 3D7 E1964 – T2291 39.6 - nt
var2CSA DBL5 3D7 K1923 – T2291 44.6 - nt
var2CSA DBL5 3D7 N1888 – T2291 48.3 + yes
var2CSA DBL6 IT/FCR3 3 E2322 – E2590 32.7 ++ yes
  1. P. pastoris clones were grown overnight at 0.9 L to build biomass and protein induced at 0.3 L scale. Yields following a single nickel chromatography step; – (not detected), + (< 1 mg/L), ++ (1 mg/L). nt, not tested.
  2. 1 Molecular weight predicted by Vector NTI software.
  3. 2 Rabbit antibodies were screened against CSA-binding infected erythrocytes and negative control A4ultra-infected erythrocytes expressing a non-VAR2CSA protein
  4. 3 Data previously published by [18].
  5. 4 Antibody reactivity could be revealed by pre-treating IEs with chymotrypsin or trypsin