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Therapeutic Enzymes: Function and Clinical Implications pp 25–54Cite as

Production of Therapeutic Enzymes by Lentivirus Transgenesis

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Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1148))

Abstract

Since ERT for several LSDs treatment has emerged at the beginning of the 1980s with Orphan Drug approval, patients’ expectancy and life quality have been improved. Most LSDs treatment are based on the replaced of mutated or deficient protein with the natural or recombinant protein.

One of the main ERT drawback is the high drug prices. Therefore, different strategies trying to optimize the global ERT biotherapeutic production have been proposed. LVs, a gene delivery tool, can be proposed as an alternative method to generate stable cell lines in manufacturing of recombinant proteins. Since LVs have been used in human gene therapy, clinical trials, safety testing assays and procedures have been developed. Moreover, one of the main advantages of LVs strategy to obtain manufacturing cell line is the short period required as well as the high protein levels achieved.

In this chapter, we will focus on LVs as a recombinant protein production platform and we will present a case study that employs LVs to express in a manufacturing cell line, alpha-Galactosidase A (rhαGAL), which is used as ERT for Fabry disease treatment.

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Abbreviations

E. coli :

Escherichia coli

ER:

endothelial reticulum

PEG:

polyethylene glycol

FDA:

Food and Drug Administration

LSD:

lysosomal storage disease

MPS:

mucopolysaccaridosis

ERT:

enzyme replacement therapy

LVs:

lentiviral vectors

CHO:

Chinese Hamster Ovary

Neu5Gc:

N-glycolylneuraminic acid

BHK-21:

baby hamster kidney cells

HEK293:

Human Embryonic Kidney 293

EMA:

European Medicines Agency

CAP:

CEVEC´s Amniocyte Production

ICH:

International Conference of Harmonization

TGE:

transient gene expression

DHFR:

dihydrofolate reductase

GS:

glutamine synthetase

MTX:

methotrexate

MSX:

methionine sulfoximine

HPRT:

hypoxanthine phosphoribosyl transferase

RMCE:

recombinase-mediated cassette exchange

FRT:

flippase recognition target sites

ZFNs:

zinc finger nucleases

TALENs:

transcription activator-like effector nucleases

NHEJ:

non-homologous end-joining

HDR:

homology-directed repair

CRISPR:

clustered regularly interspaced short palindromic repeats

SpCas9:

Streptococcus pyogenes Cas9 endonuclease

gRNA:

guide RNA

trcRNA:

trans-acting antisense RNA

PAM:

protospacer adjacent motif

S/MARs:

matrix attachment regions

UCOEs:

Ubiquitous Chromatin Opening Elements

PEI:

Polyethylenimine

RT:

reverse transcriptase

IN:

integrase

TG:

transgene

VSV:

Vesicular Stomatitis Virus

LTR:

long terminal repeats

SIN:

self-inactivating

RRE:

rev response elements

cPPT:

Central Polypurine tract

WPRE:

post-transcriptional regulatory element of the woodchuck

qPCR:

real time polymerase chain reaction

MOI:

multiplicity of infection

CAR:

chimeric antigen receptor

RCL:

replication-competent lentivirus

ELISA:

enzyme-linked immunosorbent assay

PERT:

product-enhanced reverse transcriptase

LOD:

limit of detection

M6P:

phosphate-6-O-mannose

GlcNAc:

N-acetylglucosamine

M6PR:

M6P receptor

C6S:

chondroitin-6-sulfate

KS:

keratan sulfate

GALNS:

N-acetylgalactosamine-6-sulfate sulfatase

LAL:

Lysosomal acid lipase

Gb3:

globotriaosylceramide

rhαGAL:

recombinant human alpha galactosidase A

LP:

lentiviral particle

IEX:

ionic exchange

HIC:

Hydrophobic interaction chromatography

RP-HPLC:

reversed phase high performance liquid chromatography

IEF:

isoelectrofocusing

HPAEC-PAD:

high-pH anion-exchange chromatography with pulsed amperometric detection

WAX:

weak anion exchange

Man:

Mannose

Gal:

Galactose

GlcNAc:

N-acetylglucosamine

Fuc:

Fucose

4MU-α-Gal:

4-Methylumbelliferyl α-D-galactopyranoside

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Authors and Affiliations

  1. Cell Culture Laboratory, UNL, CONICET, FBCB, Santa Fe, Argentina

    María Celeste Rodríguez, Natalia Ceaglio & Marina Etcheverrigaray

  2. Zelltek S.A., Santa Fe, Argentina

    Sebastián Antuña & María Belén Tardivo

  3. Cell Culture Laboratory, UNL, FBCB, Santa Fe, Argentina

    Claudio Prieto

Authors
  1. María Celeste Rodríguez
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  2. Natalia Ceaglio
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  3. Sebastián Antuña
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  4. María Belén Tardivo
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  5. Marina Etcheverrigaray
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  6. Claudio Prieto
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Correspondence to Claudio Prieto .

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Editors and Affiliations

  1. Department of Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos Labrou

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Rodríguez, M.C., Ceaglio, N., Antuña, S., Tardivo, M.B., Etcheverrigaray, M., Prieto, C. (2019). Production of Therapeutic Enzymes by Lentivirus Transgenesis. In: Labrou, N. (eds) Therapeutic Enzymes: Function and Clinical Implications. Advances in Experimental Medicine and Biology, vol 1148. Springer, Singapore. https://doi.org/10.1007/978-981-13-7709-9_2

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