PMID- 29034433
OWN - NLM
STAT- MEDLINE
DCOM- 20180718
LR  - 20180718
IS  - 1432-0614 (Electronic)
IS  - 0175-7598 (Linking)
VI  - 101
IP  - 22
DP  - 2017 Nov
TI  - Development of humanized scFv antibody fragment(s) that targets and blocks 
      specific HLA alleles linked to myasthenia gravis.
PG  - 8165-8179
LID - 10.1007/s00253-017-8557-1 [doi]
AB  - Myasthenia gravis (MG) is an autoimmune disease caused by sensitization of the 
      immune system to self-antigens. We have previously shown that targeting 
      MG-susceptible alleles can significantly inhibit proliferation of 
      disease-specific T cells. In this work, we humanized a murine monoclonal antibody 
      (mAb) LG11, capable of blocking MG-associated DQ beta 1 (DQB1) allele and 
      reformatted it into single-chain fragment variable (scFv). A fully functional 
      humanized scFv was obtained by optimizing variable domain orientations and linker 
      lengths, along with the optimization of expression conditions and codons to suit 
      Escherichia coli expression machinery. Characterization of humanized scFv (FL8) 
      revealed that the reformatted scFv, despite recognizing the same epitope as the 
      parent murine LG11 mAb, exhibited superior binding affinity (0.97 nM) compared to 
      the LG11 mAb, towards the immunizing antigen (DQB1*0601/70-90) and was able to 
      block the proliferation of T cells cultured from PBLs of MG-patients typed 
      DQB1*0601. The scFv was also capable of binding a variant MG-associated allele 
      (DQB1*0502/70-90) with moderate affinity (18.7 nM), a feature that was absent in 
      the LG11. To our knowledge, this is the first report of humanizing a 
      MG-associated human leukocyte antigen (HLA) scFv for preclinical studies.
FAU - Ayyar, B Vijayalakshmi
AU  - Ayyar BV
AUID- ORCID: 0000-0001-6775-9546
AD  - Department of Biochemistry and Molecular Biology, Baylor College of Medicine, 
      Houston, TX, 77030, USA. vijiayyar@gmail.com.
AD  - Department of Molecular Virology and Microbiology, Baylor College of Medicine, 
      Houston, TX, 77030, USA. vijiayyar@gmail.com.
FAU - Atassi, M Zouhair
AU  - Atassi MZ
AD  - Department of Biochemistry and Molecular Biology, Baylor College of Medicine, 
      Houston, TX, 77030, USA. matassi@bcm.edu.
AD  - Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX, 
      77030, USA. matassi@bcm.edu.
LA  - eng
PT  - Journal Article
DEP - 20171015
PL  - Germany
TA  - Appl Microbiol Biotechnol
JT  - Applied microbiology and biotechnology
JID - 8406612
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antibodies, Monoclonal, Humanized)
RN  - 0 (HLA Antigens)
RN  - 0 (Single-Chain Antibodies)
SB  - IM
MH  - *Alleles
MH  - Animals
MH  - Antibodies, Monoclonal/genetics/immunology/therapeutic use
MH  - Antibodies, Monoclonal, Humanized/genetics/*immunology/isolation & 
      purification/therapeutic use
MH  - Escherichia coli/genetics
MH  - HLA Antigens/*genetics/immunology
MH  - Humans
MH  - Mice
MH  - Myasthenia Gravis/drug therapy/genetics/*immunology
MH  - Single-Chain Antibodies/genetics/*immunology/therapeutic use
MH  - Young Adult
OTO - NOTNLM
OT  - Antibody engineering
OT  - Antibody humanization
OT  - Biacore
OT  - Expression optimization
OT  - Myasthenia gravis
OT  - scFv
EDAT- 2017/10/17 06:00
MHDA- 2018/07/19 06:00
CRDT- 2017/10/17 06:00
PHST- 2017/08/15 00:00 [received]
PHST- 2017/09/27 00:00 [accepted]
PHST- 2017/09/25 00:00 [revised]
PHST- 2017/10/17 06:00 [pubmed]
PHST- 2018/07/19 06:00 [medline]
PHST- 2017/10/17 06:00 [entrez]
AID - 10.1007/s00253-017-8557-1 [pii]
AID - 10.1007/s00253-017-8557-1 [doi]
PST - ppublish
SO  - Appl Microbiol Biotechnol. 2017 Nov;101(22):8165-8179. doi: 
      10.1007/s00253-017-8557-1. Epub 2017 Oct 15.