Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection: implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia

Udo zur Stadt; Conny Eckert; Johannes Rischewski; Katharina Michael; Steffi Golta; Manuela Müller; Reinhard Schneppenheim; Hartmut Kabisch

Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection

Standard

Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection : implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia. / zur Stadt, Udo; Eckert, Conny; Rischewski, Johannes; Michael, Katharina; Golta, Steffi; Müller, Manuela; Schneppenheim, Reinhard; Kabisch, Hartmut.

In: J CHROMATOGR B, Vol. 792, No. 2, 25.07.2003, p. 287-98.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

zur Stadt, U, Eckert, C, Rischewski, J, Michael, K, Golta, S, Müller, M, Schneppenheim, R & Kabisch, H 2003, 'Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection: implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia', J CHROMATOGR B, vol. 792, no. 2, pp. 287-98.

APA

zur Stadt, U., Eckert, C., Rischewski, J., Michael, K., Golta, S., Müller, M., Schneppenheim, R., & Kabisch, H. (2003). Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection: implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia. J CHROMATOGR B, 792(2), 287-98.

Vancouver

zur Stadt U, Eckert C, Rischewski J, Michael K, Golta S, Müller M et al. Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection: implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia. J CHROMATOGR B. 2003 Jul 25;792(2):287-98.

Bibtex

@article{278f8a8c48cb4859948e3827db257e7b,

title = "Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection: implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia",

abstract = "Incomplete T-cell-receptor delta (TCR-delta) rearrangements are widely used for detection of minimal residual disease in childhood acute lymphoblastic leukemia. In a substantial number of cases both alleles are rearranged and polymerase chain reaction (PCR) products have either to be cloned or excised and reamplified from acrylamide gels. Here we describe a novel HPLC-based method for identification and characterization of clonal TCR-delta targets. Clonality of PCR amplified TCR-delta products was examined on a high-resolution micropellicular DNASep matrix (WAVE Nucleic Acid Fragment Analysis System, Transgenomic) and subsequently classified into clonal, biclonal or negative samples. Vdelta2-Ddelta3 and Ddelta2-Ddelta3 rearrangements were analyzed by denaturing high-performance liquid chromatography (DHPLC), using triethylammonium acetate as an ion-pairing reagent, with a linear acetonitrile gradient at 50 degrees C. Biclonal elution profiles consisted of two individual homoduplex peaks and one heteroduplex peak unique for each patient sample. For characterization of biclonal rearrangements DHPLC separated samples were subjected to a second run and individual clonal peaks were collected. A software-controlled fragment collector was arranged in tandem with the HPLC system for this purpose and purified PCR products were collected in a time-dependent manner. Fractions were air dried and subsequently sequenced directly. The specificity of the observed patient specific sequences was tested via real time quantitative PCR on a LightCycler system.",

keywords = "Base Sequence, Child, Chromatography, High Pressure Liquid, DNA Probes, Gene Rearrangement, delta-Chain T-Cell Antigen Receptor, Humans, Monitoring, Physiologic, Neoplasm, Residual, Polymerase Chain Reaction, Precursor Cell Lymphoblastic Leukemia-Lymphoma, Protein Denaturation",

author = "{zur Stadt}, Udo and Conny Eckert and Johannes Rischewski and Katharina Michael and Steffi Golta and Manuela M{\"u}ller and Reinhard Schneppenheim and Hartmut Kabisch",

year = "2003",

month = jul,

day = "25",

language = "English",

volume = "792",

pages = "287--98",

journal = "J CHROMATOGR B",

issn = "1570-0232",

publisher = "Elsevier",

number = "2",

}

RIS

TY - JOUR

T1 - Identification and characterisation of clonal incomplete T-cell-receptor Vdelta2-Ddelta3/Ddelta2-Ddelta3 rearrangements by denaturing high-performance liquid chromatography and subsequent fragment collection

T2 - implications for minimal residual disease monitoring in childhood acute lymphoblastic leukemia

AU - zur Stadt, Udo

AU - Eckert, Conny

AU - Rischewski, Johannes

AU - Michael, Katharina

AU - Golta, Steffi

AU - Müller, Manuela

AU - Schneppenheim, Reinhard

AU - Kabisch, Hartmut

PY - 2003/7/25

Y1 - 2003/7/25

N2 - Incomplete T-cell-receptor delta (TCR-delta) rearrangements are widely used for detection of minimal residual disease in childhood acute lymphoblastic leukemia. In a substantial number of cases both alleles are rearranged and polymerase chain reaction (PCR) products have either to be cloned or excised and reamplified from acrylamide gels. Here we describe a novel HPLC-based method for identification and characterization of clonal TCR-delta targets. Clonality of PCR amplified TCR-delta products was examined on a high-resolution micropellicular DNASep matrix (WAVE Nucleic Acid Fragment Analysis System, Transgenomic) and subsequently classified into clonal, biclonal or negative samples. Vdelta2-Ddelta3 and Ddelta2-Ddelta3 rearrangements were analyzed by denaturing high-performance liquid chromatography (DHPLC), using triethylammonium acetate as an ion-pairing reagent, with a linear acetonitrile gradient at 50 degrees C. Biclonal elution profiles consisted of two individual homoduplex peaks and one heteroduplex peak unique for each patient sample. For characterization of biclonal rearrangements DHPLC separated samples were subjected to a second run and individual clonal peaks were collected. A software-controlled fragment collector was arranged in tandem with the HPLC system for this purpose and purified PCR products were collected in a time-dependent manner. Fractions were air dried and subsequently sequenced directly. The specificity of the observed patient specific sequences was tested via real time quantitative PCR on a LightCycler system.

AB - Incomplete T-cell-receptor delta (TCR-delta) rearrangements are widely used for detection of minimal residual disease in childhood acute lymphoblastic leukemia. In a substantial number of cases both alleles are rearranged and polymerase chain reaction (PCR) products have either to be cloned or excised and reamplified from acrylamide gels. Here we describe a novel HPLC-based method for identification and characterization of clonal TCR-delta targets. Clonality of PCR amplified TCR-delta products was examined on a high-resolution micropellicular DNASep matrix (WAVE Nucleic Acid Fragment Analysis System, Transgenomic) and subsequently classified into clonal, biclonal or negative samples. Vdelta2-Ddelta3 and Ddelta2-Ddelta3 rearrangements were analyzed by denaturing high-performance liquid chromatography (DHPLC), using triethylammonium acetate as an ion-pairing reagent, with a linear acetonitrile gradient at 50 degrees C. Biclonal elution profiles consisted of two individual homoduplex peaks and one heteroduplex peak unique for each patient sample. For characterization of biclonal rearrangements DHPLC separated samples were subjected to a second run and individual clonal peaks were collected. A software-controlled fragment collector was arranged in tandem with the HPLC system for this purpose and purified PCR products were collected in a time-dependent manner. Fractions were air dried and subsequently sequenced directly. The specificity of the observed patient specific sequences was tested via real time quantitative PCR on a LightCycler system.

KW - Base Sequence

KW - Child

KW - Chromatography, High Pressure Liquid

KW - DNA Probes

KW - Gene Rearrangement, delta-Chain T-Cell Antigen Receptor

KW - Humans

KW - Monitoring, Physiologic

KW - Neoplasm, Residual

KW - Polymerase Chain Reaction

KW - Precursor Cell Lymphoblastic Leukemia-Lymphoma

KW - Protein Denaturation

M3 - SCORING: Journal article

C2 - 12860036

VL - 792

SP - 287

EP - 298

JO - J CHROMATOGR B

JF - J CHROMATOGR B

SN - 1570-0232

IS - 2

ER -