A new purification method for the Fab and F(ab')₂ fragment of 145-2C11, hamster anti-mouse CD3ε antibody

Kwack,KyuBum The Korea Science and Technology Center 2000 BMB Reports Vol.33 No.2

Recombinant protein G has been utilized in the purification of antibodies from various mammalian species based on the interaction of antibodies with protein G. The interaction between immunoglobulin and protein G may not be restricted to the Fc portion of antibodies, as many different F(ab`)₂or Fab fragments can also bind to protein G. I found both Fab and F(ab`)₂of 145-2C11, a hamster anti-mouse CD3εantibody, bound to the protein G-sepharose. Interestingly, Fab and F(ab`)₂of 145-2C11 did not bind to the protein A-sepharose. The binding of Fab and F(ab`)₂of 145-2C11 to protein G provided a useful method to remove proteases, chopped fragments of the Fc region, and other contaminating proteins. The remaining intact antibody in the protease reaction mixture can be removed by using a protein A-sepharose, because the Fab and F(ab`)₂portions of 145-2C11 did not bind to protein A-sepharose. The specific binding of Fab and F(ab`)₂portions of 145-2C11 to a protein G-sepharose (though not to a protein A-sepharose) and binding of intact 145-2C11 to both protein A- and G-sepharose will be useful in developing an effective purification protocol for Fab and F(ab`)₂portions of 145-2C11.

Sex-specific differences in the association of a common aldehyde dehydrogenase 2 gene polymorphism and alcohol consumption with stroke risk in a Korean population

Chol Shin,KyuBum Kwack,Nam H Cho,Seong Hwan Kim,Inkyung Baik 대한지역사회영양학회 2015 Nutrition Research and Practice Vol.9 No.1

BACKGROUND/OBJECTIVES: It is well-known that alcohol consumption is associated with stroke risk as well as with aldehyde dehydrogenase 2 gene (ALDH2) polymorphisms. However, it is unclear whether ALDH2 polymorphisms are associated with stroke risk independent of alcohol consumption and whether such association is modified by sex. We evaluated sex-specific associations of a common ALDH2 polymorphism and alcohol consumption with stroke risk in a Korean population. SUBJECTS/METHODS: We conducted a prospective cohort study involving 8,465 men and women, aged 40-69 years and free of stroke between June, 2001 and January, 2003, and followed for the development of stroke. We identified new cases of stroke, which were self-reported or ascertained from vital registration data. Based on genome-wide association data, we selected a single-nucleotide polymorphism (rs2074356), which shows high linkage disequilibrium with the functional polymorphism of ALDH2. We conducted Cox proportional hazards regression analysis considering potential risk factors collected from a baseline questionnaire. RESULTS: Over the median follow-up of 8 years, 121 cases of stroke were identified. Carrying the wild-type allele of the ALDH2 polymorphism increased stroke risk among men. The multivariate hazard ratio [95% confidence interval] of stroke was 2.02 [1.03-3.99] for the wild-type allele compared with the mutant alleles, but the association was attenuated after controlling for alcohol consumption. Combinations of the wild-type allele and other risk factors of stroke, such as old age, diabetes mellitus, and habitual snoring, synergistically increased the risk among men. Among women, however, the ALDH2 polymorphism was not associated with stroke risk. CONCLUSIONS: The prospective cohort study showed a significant association between a common ALDH2 polymorphism and stroke risk in Korean men, but not in Korean women, and also demonstrated that men with genetic disadvantages gain more risk when having risk factors of stroke. Thus, these men may need to make more concerted efforts to control modifiable risk factors of stroke.

CD137 induces adhesion and cytokine production in human monocytic THP-1cells

최정원,이현우,노구섭,김홍희,KyuBum Kwack 생화학분자생물학회 2005 Experimental and molecular medicine Vol.37 No.2

CD137, which is expressed on activated T cells, plays a critical role in inflammatory responses. However, the exact role that CD137 plays in monocytes is not fully known. Here we studied the expression and function of CD137 in human monocytic THP-1 cells, which we found constitutively expresses CD137 at the mRNA and protein level. Cross-linking of CD137 increased the secretion of IL-8 and TNF-a, promoted the expression of CD54 and CD11b, and increased adhesion to ECM proteins. In particular CD137-induced adhesion of THP-cells was inhibited by an inhibitor of MEK, but not by a p38 kinase inhibitor. Taken together, these results show that the adhesion and cytokine production of THP-1 cells induced by CD137 occurs via activation of MEK, which results in the activation of ERK-1/2 signaling pathways. Therefore, this study suggests that CD137 induces an activating and migrating signal during inflammatory processes.

Interaction between thyroglobulin and ADAMTS16 in premature ovarian failure

Pyun, Jung-A,Kim, Sunshin,Kwack, KyuBum The Korean Society for Reproductive Medicine 2014 Clinical and Experimental Reproductive Medicine Vol.41 No.3

Objective: The aim of the present study was to examine whether interactions between polymorphisms in the thyroglobulin and ADAM metallopeptidase with thrombospondin type 1 motif, 16 (ADAMTS16) genes are associated with the development of premature ovarian failure (POF). Methods: A total of 75 patients with POF and 196 controls were involved in this study. We used a GoldenGate assay to genotype single nucleotide polymorphisms (SNPs). Logistic regression analysis was performed to identify POF-associated polymorphisms and synergistic interactions between polymorphisms in the thyroglobulin and ADAMTS16 genes. Results: Single gene analyses using logistic regression analysis showed no significant association between polymorphisms in the two genes and POF. In the results from interaction analyses, we found seven synergistic interactions between the polymorphisms in thyroglobulin and ADAMTS16, although there was no combination showing p-values lower than the significant threshold using the Bonferroni correction. When the AG genotype was present at the rs853326 missense SNP, the A and G alleles at the tagging SNPs rs16875268 and rs13168665 showed significant interactions (odds ratios=5.318 and 16.2 respectively; 95% confidence intervals, 1.64-17.28 and 2.08-126.4; p=0.0054 and 0.0079). Conclusion: Synergistic interactions between polymorphisms in the thyroglobulin and ADAMTS16 genes were associated with an increased risk of POF development in Korean women.

Epistasis between polymorphisms in ACVR2B and ADAMTS19 is associated with premature ovarian failure

Pyun, Jung-A,Kim, Sunshin,Kwack, KyuBum by The North American Menopause Society. 2015 Menopause Vol.22 No.2

OBJECTIVE: This study investigated whether epistasis between single nucleotide polymorphisms (SNPs) within ACVR2B (activin A receptor, type IIB) and ADAMTS19 (ADAM metallopeptidase with thrombospondin type 1 motif, 19) genes is associated with premature ovarian failure (POF). METHODS: One hundred twenty women with POF and 152 controls were recruited for stage I, and 1,641 additional female controls participated in stage II. GoldenGate assay with VeraCode technology was used for genotyping ACVR2B and ADAMTS19 SNPs in stage I. In stage II, we obtained genotype data for SNPs using Affymetrix Genome-Wide Human SNP array 5.0 and imputed data using IMPUTE program from the Korean Genome Epidemiology Study. RESULTS: In stage I, five combinations showed significant synergistic interactions after Bonferroni correction. One SNP (rs1468077 within 5&vprime; flanking region) and two intronic SNPs (rs2268753 and rs2268757) in ACVR2B and three intronic SNPs within ADAMTS19 (rs13158524, rs1476083, and rs1972624) were involved in synergistic interactions in a recessive manner. In stage II and combined analyses, we could not find any significant interactions between the SNPs. However, diplotypes within ACVR2B and ADAMTS19 that consist of risk genotypes or alleles in the results of significant synergistic interactions between SNPs showed significant interactions after Bonferroni correction. Thirteen and nine significant synergistic interactions were found in a dominant model in stage II and combined analyses, respectively (strongest association in combined analysis: odds ratio, 5.93; 95% CI, 2.47-14.20; P = 6.65 × 10). CONCLUSIONS: Epistasis between polymorphisms within ACVR2B and ADAMTS19 is significantly associated with susceptibility to POF.

Epistasis between polymorphisms in PCSK1 and DBH is associated with premature ovarian failure

Pyun, Jung-A,Kim, Sunshin,Cha, Dong Hyun,Kwack, KyuBum by The North American Menopause Society. 2014 Menopause Vol.21 No.11

OBJECTIVE: This study examined whether epistasis between single nucleotide polymorphisms (SNPs) within proprotein convertase subtilisin/kexin type 1 (PCSK1) and dopamine &bgr;-hydroxylase (DBH) genes is associated with premature ovarian failure (POF). METHODS: One hundred twenty women with POF and 222 female controls were recruited for this study. To genotype SNPs within PCSK1 and DBH, we used a GoldenGate assay with VeraCode technology, which uses an allele-specific primer extension method. RESULTS: Two SNPs (rs155979 and rs3762986) within PCSK1 and one SNP (rs1611114) within DBH, which were located in the 5&vprime; flanking region, were involved in synergistic interactions. The C allele in the rs155979 SNP showed an increased risk of POF in a dominant model when AA genotype in the rs1611114 SNP was present (odds ratio, 3.60; 95% CI, 1.82-7.14; P = 0.00024), whereas the G allele in the rs1611114 SNP showed a reduced risk of POF in a dominant model when at least one C allele at the rs155979 SNP was present (odds ratio, 0.24; 95% CI, 0.11-0.51; P = 0.00018) or one G allele at the rs3762986 SNP was present (odds ratio, 0.33; 95% CI, 0.19-0.60; P = 0.00023). CONCLUSIONS: Epistases between SNPs within PCSK1 and DBH genes are significantly associated with susceptibility or resistance to POF.

Association of <i>GNLY</i> Genetic Polymorphisms with Chronic Liver Disease in a Korean Population

Park, Geun-Hee,Kim, Kyoung-Yeon,Cheong, Jae Youn,Cho, Sung Won,Kwack, KyuBum Mary Ann Liebert 2012 DNA and cell biology Vol.31 No.9

Lymph Node Metastasis of Gastric Cancer Is Associated with the Interaction Between Poly (ADP-Ribose) Polymerase 1 and Matrix Metallopeptidase 2

Kim, JiHye,Pyun, Jung-A,Won Cho, Sung,Lee, KwangJae,Kwack, KyuBum Mary Ann Liebert 2011 DNA and cell biology Vol.30 No.12

<P>Poly (ADP-ribose) polymerase 1 (PARP1), which plays a critical role in the base excision DNA repair mechanism, and matrix metallopeptidase 2 (MMP2), a member of the matrix metalloprotease family, are involved in tumor formation and metastasis, respectively. In the present study, the possible association of single nucleotide polymorphisms (SNPs) and gene-gene interaction between PARP1 and MMP2 with the increased incidence of gastric cancer (GC) development and lymph node metastasis (LNM) was investigated in a Korean population. Samples were obtained from 326 patients with chronic gastritis and 153 patients with GC and genotyped using the GoldenGate? method. The PARP1 rs1136410 genotype showed a significant association with the frequency of LNM of GC (odds ratio [OR]?=?2.19, p?=?0.02), LNM stage (p?=?0.035), and tumor invasion (p?=?0.035). The allele frequency of MMP2 rs243865 was not associated with the development of GC or with the development of LNM of GC. Epistasis between the PARP1 SNP and the MMP2 SNP was associated with the development of LNM of GC. The combination of the MMP2 rs243865 CC genotype and the PARP1 rs1136410 CC or CC+CT genotypes showed a high risk of LNM of GC (OR?=?2.47, p?=?0.01; OR?=?2.28, p?=?0.01, respectively). In summary, PARP1 is associated with the risk of LNM of GC and the stage of LNM and tumor invasion. Epistasis between PARP1 rs1136410 and MMP2 rs243865 increased the risk of LNM of GC.</P>

Polymorphisms within the FANCA gene associate with premature ovarian failure in Korean women

Pyun, Jung-A.,Kim, Sunshin,Cha, Dong Hyun,Kwack, KyuBum by The North American Menopause Society. 2014 Menopause Vol.21 No.5

OBJECTIVE: This study investigated whether polymorphisms within the Fanconi anemia complementation group A (FANCA) gene contribute to the increased risk of premature ovarian failure (POF) in Korean women. METHODS: Ninety-eight women with POF and 218 controls participated in this study. Genomic DNA from peripheral blood was isolated, and GoldenGate genotyping assay was used to identify single nucleotide polymorphisms (SNPs) within the FANCA gene. RESULTS: Two significant SNPs (rs1006547 and rs2239359; P < 0.05) were identified by logistic regression analysis, but results were insignificant after Bonferroni correction. Six SNPs formed a linkage disequilibrium block, and three main haplotypes were found. Two of three haplotypes (AAAGAA and GGGAGG) distributed highly in the POF group, whereas the remaining haplotype (GGAAGG) distributed highly in the control group by logistic regression analysis (highest odds ratio, 2.515; 95% CI, 1.515-4.175; P = 0.00036). CONCLUSIONS: Our observations suggest that genetic variations in the FANCA gene may increase the risk for POF in Korean women.

Epistasis between polymorphisms in TSHB and ADAMTS16 is associated with premature ovarian failure

Pyun, Jung-A.,Kim, Sunshin,Cha, Dong Hyun,Kwack, KyuBum by The North American Menopause Society. 2014 Menopause Vol.21 No.8

OBJECTIVE: This study investigated whether epistasis between single nucleotide polymorphisms (SNPs) within the TSHB (thyroid-stimulating hormone &bgr;) and ADAMTS16 (ADAM metallopeptidase with thrombospondin type 1 motif, 16) genes is associated with an increased risk of premature ovarian failure (POF) in Korean women. METHODS: In stage I, 120 women with POF and 222 controls participated. A GoldenGate assay with VeraCode technology was used to genotype SNPs within the TSHB and ADAMTS16 genes. For stage II, we obtained genotype data merged with imputed data for 1,641 female controls from the Korean Genome Epidemiology Study. RESULTS: In stage I, two SNPs (rs7530810 and rs1321108) in the 5&vprime; flanking region of the TSHB gene demonstrated significant synergistic interactions with one tagging intronic SNP (rs13172105) in the ADAMTS16 gene (odds ratios, 6.63 and 5.57; 95% CIs, 2.30-19.18 and 2.05-15.12; P = 0.00048 and 0.00074, respectively) although the SNPs were not significantly associated with POF in a single SNP model. When at least one G allele at rs7530810 or one A allele at rs1321108 was present in combination with a C allele at rs13172105, significant synergistic effects were observed in a recessive model. In stage II and combined analyses, the same combinations repeatedly showed significant synergistic interactions. CONCLUSIONS: Epistasis between SNPs within the TSHB and ADAMTS16 genes may increase the risk of POF in Korean women.