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- PDB-5vh3: Crystal structure of Fab fragment of the anti-TNFa antibody infli... -

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Basic information

Entry
Database: PDB / ID: 5vh3
TitleCrystal structure of Fab fragment of the anti-TNFa antibody infliximab in a C-centered orthorhombic crystal form
Components
  • Infliximab Fab Heavy Chain
  • Infliximab Fab Light Chain
KeywordsIMMUNE SYSTEM / antibody / Fab / infliximab / biosimilar / TNFa
Function / homologyImmunoglobulins / Immunoglobulin-like / Sandwich / Mainly Beta
Function and homology information
Biological speciesMus musculus (house mouse)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2 Å
AuthorsMayclin, S.J. / Edwards, T.E. / Lerch, T.F. / Conlan, H. / Sharpe, P.
CitationJournal: MAbs / Year: 2017
Title: Infliximab crystal structures reveal insights into self-association.
Authors: Lerch, T.F. / Sharpe, P. / Mayclin, S.J. / Edwards, T.E. / Lee, E. / Conlon, H.D. / Polleck, S. / Rouse, J.C. / Luo, Y. / Zou, Q.
History
DepositionApr 12, 2017Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 3, 2017Provider: repository / Type: Initial release
Revision 1.1Jun 7, 2017Group: Database references
Revision 1.2Aug 2, 2017Group: Database references / Category: citation
Item: _citation.journal_volume / _citation.page_first / _citation.page_last
Revision 1.3Oct 4, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
H: Infliximab Fab Heavy Chain
L: Infliximab Fab Light Chain
A: Infliximab Fab Heavy Chain
B: Infliximab Fab Light Chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)95,6965
Polymers95,6344
Non-polymers621
Water18,8441046
1
H: Infliximab Fab Heavy Chain
L: Infliximab Fab Light Chain


Theoretical massNumber of molelcules
Total (without water)47,8172
Polymers47,8172
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3170 Å2
ΔGint-26 kcal/mol
Surface area19640 Å2
MethodPISA
2
A: Infliximab Fab Heavy Chain
B: Infliximab Fab Light Chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)47,8793
Polymers47,8172
Non-polymers621
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3470 Å2
ΔGint-22 kcal/mol
Surface area19590 Å2
MethodPISA
Unit cell
Length a, b, c (Å)86.580, 138.820, 195.460
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number20
Space group name H-MC2221

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Components

#1: Antibody Infliximab Fab Heavy Chain


Mass: 24356.178 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Production host: Cricetulus griseus (Chinese hamster)
#2: Antibody Infliximab Fab Light Chain


Mass: 23460.873 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Production host: Cricetulus griseus (Chinese hamster)
#3: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C2H6O2
#4: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 1046 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 3.07 Å3/Da / Density % sol: 59.94 %
Crystal growTemperature: 289 K / Method: vapor diffusion, sitting drop / pH: 7.5
Details: Fab at 10 mg/mL against Wiz 34 screen condition B4 20% PEG 3350, 0.2 M potassium citrate tribasic supplemented with 20% ethylene glycol as cryo-protectant, crystal tracking ID 267664b4, unique puck ID sdw5-2

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-G / Wavelength: 0.97856 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Nov 19, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97856 Å / Relative weight: 1
ReflectionResolution: 2→50 Å / Num. obs: 79590 / % possible obs: 100 % / Observed criterion σ(I): -3 / Redundancy: 6.246 % / Biso Wilson estimate: 21.24 Å2 / CC1/2: 0.994 / Rmerge(I) obs: 0.126 / Rrim(I) all: 0.137 / Χ2: 0.964 / Net I/σ(I): 10.52
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2Rrim(I) all% possible all
2-2.056.2890.5393.758300.8890.588100
2.05-2.116.2910.4584.2957020.9080.5100
2.11-2.176.2970.385.0155210.9390.414100
2.17-2.246.3080.3245.8253610.9490.353100
2.24-2.316.2960.2876.4152200.9590.313100
2.31-2.396.3110.2666.8550490.9630.29100
2.39-2.486.2860.2257.7648800.9730.246100
2.48-2.586.2920.1958.8146880.9780.212100
2.58-2.76.2830.16510.1444980.9840.18100
2.7-2.836.2650.14411.2743430.9870.157100
2.83-2.986.2650.12412.9140810.9890.135100
2.98-3.166.2320.10714.739050.9910.116100
3.16-3.386.2140.09416.3636450.9910.10399.9
3.38-3.656.1650.08517.7334440.9930.09399.9
3.65-46.1830.08218.6631560.9920.0999.9
4-4.476.1810.07819.4628660.9920.085100
4.47-5.166.1620.07520.1325490.9930.082100
5.16-6.326.1020.07519.2521820.9930.082100
6.32-8.946.0010.07219.3716990.9920.07999.9
8.94-505.5540.06619.819710.9920.07397.9

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
XSCALEdata scaling
Cootmodel building
PHENIXdev_2229refinement
PDB_EXTRACT3.22data extraction
XDSdata reduction
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4g3y
Resolution: 2→50 Å / SU ML: 0.2 / Cross valid method: FREE R-VALUE / σ(F): 1.36 / Phase error: 19.6
RfactorNum. reflection% reflection
Rfree0.1996 3868 4.86 %
Rwork0.1597 --
obs0.1617 79576 99.95 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Displacement parametersBiso max: 91.11 Å2 / Biso mean: 27.218 Å2 / Biso min: 4.1 Å2
Refinement stepCycle: final / Resolution: 2→50 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6515 0 4 1047 7566
Biso mean--46.02 36.27 -
Num. residues----864
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0066696
X-RAY DIFFRACTIONf_angle_d0.7889130
X-RAY DIFFRACTIONf_chiral_restr0.0531028
X-RAY DIFFRACTIONf_plane_restr0.0051171
X-RAY DIFFRACTIONf_dihedral_angle_d13.083975
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 28

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
2-2.02440.27781220.224626672789100
2.0244-2.050.25721340.196827112845100
2.05-2.0770.25931460.189226462792100
2.077-2.10540.26071430.188527122855100
2.1054-2.13550.22941160.18526502766100
2.1355-2.16740.20631380.172326992837100
2.1674-2.20130.21681330.165826862819100
2.2013-2.23730.21191190.170426882807100
2.2373-2.27590.19931310.172727272858100
2.2759-2.31730.20731310.169126252756100
2.3173-2.36190.22971410.16926992840100
2.3619-2.41010.20761190.17427382857100
2.4101-2.46250.22181370.170526652802100
2.4625-2.51980.20361250.173426882813100
2.5198-2.58280.22091470.165826972844100
2.5828-2.65260.20361520.166126692821100
2.6526-2.73060.21751520.1726952847100
2.7306-2.81880.2451500.166926882838100
2.8188-2.91950.1871250.165927242849100
2.9195-3.03640.21011340.171927072841100
3.0364-3.17450.21981250.16827062831100
3.1745-3.34190.18871410.158427132854100
3.3419-3.55120.16681470.154327072854100
3.5512-3.82530.18851600.148426832843100
3.8253-4.210.18421540.131927322886100
4.21-4.81870.14321600.113227552915100
4.8187-6.06910.16931220.134527782900100
6.0691-47.51730.20621640.17282853301799
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
11.5785-0.1369-0.25732.07331.282.4326-0.0386-0.05110.1003-0.07080.00210.2479-0.0835-0.2338-0.02410.11340.0574-0.02520.15660.0280.18-32.4482-16.481114.699
20.4009-0.33160.24472.50930.55020.5365-0.0184-0.01640.0417-0.05130.03240.26010.0006-0.08420.03890.12880.05150.01420.17160.04130.1692-29.9301-21.876420.2868
32.52920.6045-1.24312.555-0.11754.40690.09020.0795-0.18370.1357-0.05130.02760.1989-0.3706-0.04690.1490.0217-0.04280.1635-0.00370.1855-21.4325-51.674325.9508
43.9707-1.10141.79623.17030.72871.3094-0.0408-0.2109-0.2820.6496-0.2042-0.02690.062-0.04280.06120.24260.08430.01060.1831-0.01680.1688-12.9422-18.928235.3305
51.57980.21670.15043.2464-0.83591.7656-0.1231-0.38710.03880.53460.14550.0923-0.0504-0.0788-0.01290.24170.1328-0.00280.2619-0.0120.1436-20.9121-11.665535.5441
61.22020.0018-0.07392.3056-1.23013.1882-0.2887-0.3572-0.00250.5190.28780.04740.027-0.0904-0.03030.22230.12960.01340.24570.02180.1505-20.7743-15.535831.9969
74.353-4.53161.57186.3437-2.52643.1311-0.03470.23240.2176-0.0709-0.1371-0.1617-0.4616-0.01780.1740.18380.012-0.02930.1131-0.00960.1205-11.4791-33.510641.9641
82.97740.35221.09343.2261.9477.30240.31140.3174-0.5549-0.222-0.15350.24830.4364-0.3889-0.11050.24610.0569-0.07310.1535-0.05070.2882-10.723-55.710322.1121
91.4821-2.27-2.93354.58675.28016.93310.20130.00910.1041-0.335-0.0665-0.108-0.29750.0292-0.09120.17390.0455-0.01880.120.01260.1241-6.2644-43.47929.3926
104.4658-3.5396-2.7952.84662.26911.77760.44110.37680.5443-0.1323-0.2896-0.2392-0.2237-0.2168-0.22290.28110.06440.00540.17870.03740.2219-2.2966-43.61520.8572
116.64330.3451-3.50614.10063.23634.73030.05140.0890.0177-0.0961-0.13270.0636-0.2813-0.67820.0470.19940.0279-0.01890.1801-0.01820.1573-17.1902-37.787138.3941
123.2954-2.1475-1.40384.15873.67475.35850.220.1708-0.1636-0.14040.0469-0.13450.170.1922-0.26790.16670.03410.00170.12920.01520.182-1.7446-51.186724.6613
130.9916-0.23590.36761.9946-1.16682.5468-0.03390.0008-0.0458-0.04630.0306-0.29340.0870.27770.00280.11320.0485-0.00350.1778-0.05050.18229.7259-52.570515.2264
141.2222-1.01760.07964.11121.14833.1137-0.0339-0.01720.2730.08350.0907-0.2994-0.39070.1837-0.05460.1425-0.0045-0.01290.1769-0.02160.224919.7278-17.970321.6193
151.3865-0.5284-0.32942.22110.35850.853-0.1773-0.24070.01710.5150.1795-0.08830.04390.16930.02870.26520.1358-0.0370.2552-0.01440.162717.7554-54.287335.0212
160.484-1.2347-1.01022.79261.93380.9417-0.1885-0.12120.05920.4030.2903-0.11040.20710.3158-0.10210.21990.091-0.03680.2164-0.03530.186316.8882-47.153634.0548
173.83-0.1661-1.33615.1689-2.32946.82210.35020.38180.6126-0.0298-0.1013-0.3099-0.93050.0481-0.12150.20230.05560.03910.13390.04310.27128.2305-12.564920.2359
182.1973-2.44023.48364.8125-5.47116.93120.12930.0227-0.108-0.1588-0.05630.03020.12150.064-0.05770.1420.0484-0.00160.1022-0.0410.11384.5581-24.240828.7027
192.627-2.14620.57062.0936-1.16942.01290.09290.2808-0.0689-0.0865-0.17530.090.01770.40150.02980.13860.0158-0.00090.0966-0.0090.16877.664-26.910228.3765
202.8598-2.13771.52754.6285-4.59536.97340.1480.17290.06880.00980.15450.1575-0.1726-0.3652-0.31590.10910.0198-0.03630.1389-0.0150.1832-0.3945-16.914723.9487
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'H' and (resid 1 through 93 )H1 - 93
2X-RAY DIFFRACTION2chain 'H' and (resid 94 through 126 )H94 - 126
3X-RAY DIFFRACTION3chain 'H' and (resid 127 through 222 )H127 - 222
4X-RAY DIFFRACTION4chain 'L' and (resid 1 through 18 )L1 - 18
5X-RAY DIFFRACTION5chain 'L' and (resid 19 through 75 )L19 - 75
6X-RAY DIFFRACTION6chain 'L' and (resid 76 through 102 )L76 - 102
7X-RAY DIFFRACTION7chain 'L' and (resid 103 through 113 )L103 - 113
8X-RAY DIFFRACTION8chain 'L' and (resid 114 through 128 )L114 - 128
9X-RAY DIFFRACTION9chain 'L' and (resid 129 through 150 )L129 - 150
10X-RAY DIFFRACTION10chain 'L' and (resid 151 through 163 )L151 - 163
11X-RAY DIFFRACTION11chain 'L' and (resid 164 through 174 )L164 - 174
12X-RAY DIFFRACTION12chain 'L' and (resid 175 through 214 )L175 - 214
13X-RAY DIFFRACTION13chain 'A' and (resid 1 through 113 )A1 - 113
14X-RAY DIFFRACTION14chain 'A' and (resid 114 through 222 )A114 - 222
15X-RAY DIFFRACTION15chain 'B' and (resid 1 through 75 )B1 - 75
16X-RAY DIFFRACTION16chain 'B' and (resid 76 through 113 )B76 - 113
17X-RAY DIFFRACTION17chain 'B' and (resid 114 through 128 )B114 - 128
18X-RAY DIFFRACTION18chain 'B' and (resid 129 through 150 )B129 - 150
19X-RAY DIFFRACTION19chain 'B' and (resid 151 through 174 )B151 - 174
20X-RAY DIFFRACTION20chain 'B' and (resid 175 through 214 )B175 - 214

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