Hexa-thiocarbamoyl phenyl PEG5K Hb: Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts

Fantao Meng; Belur N. Manjula; Amy G. Tsai; Pedro Cabrales; Marcos Intaglietta; Paul K. Smith; Muthuchidambaram Prabhakaran; Seetharama A. Acharya

doi:10.1007/s10930-009-9185-0

Hexa-thiocarbamoyl phenyl PEG5K Hb: Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts

Fantao Meng, Belur N. Manjula, Amy G. Tsai, Pedro Cabrales, Marcos Intaglietta, Paul K. Smith, Muthuchidambaram Prabhakaran, Seetharama A. Acharya

Medicine

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

A new hexaPEGylated hemoglobin, (TCP-PEG5K)₆-Hb (TCP, thiocarbamoyl phenyl) has been developed using PEG-phenyl-isothiocyanate and its vasoactivity and structure has been investigated. Of the six PEG5K chains of (TCP-PEG5K)₆-Hb, 4 are conjugated to the α-amino groups of Hb, and the other 2 chains are distributed on ε-amino groups, identified as Lys-40(α) (∼45%), Lys-56(α) (∼25%), and Lys-8(β) (∼24%). The studies with hamster infused with a bolus of a 4 gm % solution of (TCP-PEG5K)₆-Hb equivalent to 10% of their blood volume have established that this new hexaPEGylated Hb is vasoinactive. The viscosity and the colloidal osmotic pressure of (TCP-PEG5K)₆-Hb at 4% is 1.9 cP and 69.7 mmHg, respectively. The molecular radius of (TCP-PEG5K)₆-Hb is about 4.6 nm and is significantly smaller than hexaPEGylated Hbs developed using other direct and extension arm facilitated PEGylation platform. The presence of an outside the central cavity intramolecular crosslink, succinimidophenyl-PEG2K between Cys-93(β, β′) in (TCP-PEG5K)₆-ββ- Hb strongly impacts its solution properties. These patterns of influence suggest that the inter-dimeric interactions in (TCP-PEG5K)₆-Hb is weakened just as with other direct PEGylation platforms, and (SP-PEG5K)₆-Hb generated by EAF-PEGylation is unique in not inducing this effect. A comparison of the properties of hexaPEGylated Hbs establishes that rigidity of the conjugation linkage between PEG and Hb plays a significant influence on the resultant dictating solution properties and/structure/conformation of PEG-Hb adduct.

Original language	English (US)
Pages (from-to)	199-212
Number of pages	14
Journal	Protein Journal
Volume	28
Issue number	5
DOIs	https://doi.org/10.1007/s10930-009-9185-0
State	Published - Jun 2009

Keywords

Blood substitute
Conjugation chemistry
Hemoglobin
Linkage rigidity
PEGylation
Vasoactivity

ASJC Scopus subject areas

Analytical Chemistry
Bioengineering
Biochemistry
Organic Chemistry

Access to Document

10.1007/s10930-009-9185-0

Cite this

Meng, F., Manjula, B. N., Tsai, A. G., Cabrales, P., Intaglietta, M., Smith, P. K., Prabhakaran, M., & Acharya, S. A. (2009). Hexa-thiocarbamoyl phenyl PEG5K Hb: Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts. Protein Journal, 28(5), 199-212. https://doi.org/10.1007/s10930-009-9185-0

Meng, F, Manjula, BN, Tsai, AG, Cabrales, P, Intaglietta, M, Smith, PK, Prabhakaran, M & Acharya, SA 2009, 'Hexa-thiocarbamoyl phenyl PEG5K Hb: Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts', Protein Journal, vol. 28, no. 5, pp. 199-212. https://doi.org/10.1007/s10930-009-9185-0

@article{2f3b0acf1af641deba35587a149ab876,

title = "Hexa-thiocarbamoyl phenyl PEG5K Hb: Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts",

abstract = "A new hexaPEGylated hemoglobin, (TCP-PEG5K)6-Hb (TCP, thiocarbamoyl phenyl) has been developed using PEG-phenyl-isothiocyanate and its vasoactivity and structure has been investigated. Of the six PEG5K chains of (TCP-PEG5K)6-Hb, 4 are conjugated to the α-amino groups of Hb, and the other 2 chains are distributed on ε-amino groups, identified as Lys-40(α) (∼45%), Lys-56(α) (∼25%), and Lys-8(β) (∼24%). The studies with hamster infused with a bolus of a 4 gm % solution of (TCP-PEG5K)6-Hb equivalent to 10% of their blood volume have established that this new hexaPEGylated Hb is vasoinactive. The viscosity and the colloidal osmotic pressure of (TCP-PEG5K)6-Hb at 4% is 1.9 cP and 69.7 mmHg, respectively. The molecular radius of (TCP-PEG5K)6-Hb is about 4.6 nm and is significantly smaller than hexaPEGylated Hbs developed using other direct and extension arm facilitated PEGylation platform. The presence of an outside the central cavity intramolecular crosslink, succinimidophenyl-PEG2K between Cys-93(β, β′) in (TCP-PEG5K)6-ββ- Hb strongly impacts its solution properties. These patterns of influence suggest that the inter-dimeric interactions in (TCP-PEG5K)6-Hb is weakened just as with other direct PEGylation platforms, and (SP-PEG5K)6-Hb generated by EAF-PEGylation is unique in not inducing this effect. A comparison of the properties of hexaPEGylated Hbs establishes that rigidity of the conjugation linkage between PEG and Hb plays a significant influence on the resultant dictating solution properties and/structure/conformation of PEG-Hb adduct.",

keywords = "Blood substitute, Conjugation chemistry, Hemoglobin, Linkage rigidity, PEGylation, Vasoactivity",

author = "Fantao Meng and Manjula, {Belur N.} and Tsai, {Amy G.} and Pedro Cabrales and Marcos Intaglietta and Smith, {Paul K.} and Muthuchidambaram Prabhakaran and Acharya, {Seetharama A.}",

note = "Funding Information: Acknowledgments We thank Dr. Parimala Nacharaju, Dr. Tao Hu, Dr. Dongxia Li, Dr. Ananda U, and Dr. Maria Lobo for critical review of this paper prior to submission. We would like to acknowledge the assistance of Edward Nieves for the help in MALDI-TOF analysis (the Macromolecular Analysis and Proteomics Laboratory at the Albert Einstein College of Medicine). This research was supported by a grant-in-aid from the American Heart Association Heritage Affiliate, the National Institutes of Health grants HL58247, HL71064 and USPHS NIH Bioengineering Partnership grant 1R24 HL 64395, and the US Army grant PR023085.",

year = "2009",

month = jun,

doi = "10.1007/s10930-009-9185-0",

language = "English (US)",

volume = "28",

pages = "199--212",

journal = "Protein Journal",

issn = "1572-3887",

publisher = "Springer New York",

number = "5",

}

TY - JOUR

T1 - Hexa-thiocarbamoyl phenyl PEG5K Hb

T2 - Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts

AU - Meng, Fantao

AU - Manjula, Belur N.

AU - Tsai, Amy G.

AU - Cabrales, Pedro

AU - Intaglietta, Marcos

AU - Smith, Paul K.

AU - Prabhakaran, Muthuchidambaram

AU - Acharya, Seetharama A.

N1 - Funding Information: Acknowledgments We thank Dr. Parimala Nacharaju, Dr. Tao Hu, Dr. Dongxia Li, Dr. Ananda U, and Dr. Maria Lobo for critical review of this paper prior to submission. We would like to acknowledge the assistance of Edward Nieves for the help in MALDI-TOF analysis (the Macromolecular Analysis and Proteomics Laboratory at the Albert Einstein College of Medicine). This research was supported by a grant-in-aid from the American Heart Association Heritage Affiliate, the National Institutes of Health grants HL58247, HL71064 and USPHS NIH Bioengineering Partnership grant 1R24 HL 64395, and the US Army grant PR023085.

PY - 2009/6

Y1 - 2009/6

N2 - A new hexaPEGylated hemoglobin, (TCP-PEG5K)6-Hb (TCP, thiocarbamoyl phenyl) has been developed using PEG-phenyl-isothiocyanate and its vasoactivity and structure has been investigated. Of the six PEG5K chains of (TCP-PEG5K)6-Hb, 4 are conjugated to the α-amino groups of Hb, and the other 2 chains are distributed on ε-amino groups, identified as Lys-40(α) (∼45%), Lys-56(α) (∼25%), and Lys-8(β) (∼24%). The studies with hamster infused with a bolus of a 4 gm % solution of (TCP-PEG5K)6-Hb equivalent to 10% of their blood volume have established that this new hexaPEGylated Hb is vasoinactive. The viscosity and the colloidal osmotic pressure of (TCP-PEG5K)6-Hb at 4% is 1.9 cP and 69.7 mmHg, respectively. The molecular radius of (TCP-PEG5K)6-Hb is about 4.6 nm and is significantly smaller than hexaPEGylated Hbs developed using other direct and extension arm facilitated PEGylation platform. The presence of an outside the central cavity intramolecular crosslink, succinimidophenyl-PEG2K between Cys-93(β, β′) in (TCP-PEG5K)6-ββ- Hb strongly impacts its solution properties. These patterns of influence suggest that the inter-dimeric interactions in (TCP-PEG5K)6-Hb is weakened just as with other direct PEGylation platforms, and (SP-PEG5K)6-Hb generated by EAF-PEGylation is unique in not inducing this effect. A comparison of the properties of hexaPEGylated Hbs establishes that rigidity of the conjugation linkage between PEG and Hb plays a significant influence on the resultant dictating solution properties and/structure/conformation of PEG-Hb adduct.

AB - A new hexaPEGylated hemoglobin, (TCP-PEG5K)6-Hb (TCP, thiocarbamoyl phenyl) has been developed using PEG-phenyl-isothiocyanate and its vasoactivity and structure has been investigated. Of the six PEG5K chains of (TCP-PEG5K)6-Hb, 4 are conjugated to the α-amino groups of Hb, and the other 2 chains are distributed on ε-amino groups, identified as Lys-40(α) (∼45%), Lys-56(α) (∼25%), and Lys-8(β) (∼24%). The studies with hamster infused with a bolus of a 4 gm % solution of (TCP-PEG5K)6-Hb equivalent to 10% of their blood volume have established that this new hexaPEGylated Hb is vasoinactive. The viscosity and the colloidal osmotic pressure of (TCP-PEG5K)6-Hb at 4% is 1.9 cP and 69.7 mmHg, respectively. The molecular radius of (TCP-PEG5K)6-Hb is about 4.6 nm and is significantly smaller than hexaPEGylated Hbs developed using other direct and extension arm facilitated PEGylation platform. The presence of an outside the central cavity intramolecular crosslink, succinimidophenyl-PEG2K between Cys-93(β, β′) in (TCP-PEG5K)6-ββ- Hb strongly impacts its solution properties. These patterns of influence suggest that the inter-dimeric interactions in (TCP-PEG5K)6-Hb is weakened just as with other direct PEGylation platforms, and (SP-PEG5K)6-Hb generated by EAF-PEGylation is unique in not inducing this effect. A comparison of the properties of hexaPEGylated Hbs establishes that rigidity of the conjugation linkage between PEG and Hb plays a significant influence on the resultant dictating solution properties and/structure/conformation of PEG-Hb adduct.

KW - Blood substitute

KW - Conjugation chemistry

KW - Hemoglobin

KW - Linkage rigidity

KW - PEGylation

KW - Vasoactivity

UR - http://www.scopus.com/inward/record.url?scp=69549114297&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69549114297&partnerID=8YFLogxK

U2 - 10.1007/s10930-009-9185-0

DO - 10.1007/s10930-009-9185-0

M3 - Article

C2 - 19653083

AN - SCOPUS:69549114297

SN - 1572-3887

VL - 28

SP - 199

EP - 212

JO - Protein Journal

JF - Protein Journal

IS - 5

ER -

Hexa-thiocarbamoyl phenyl PEG5K Hb: Vasoactivity and structure : Ithe conjugation linkage on the pegylation induced plasma expander-like solution properties of PEG-Hb adducts

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this