A5014389802- Monoclonal and Polyclonal Antibodies Research
- Chemical Synthesis and Analysis
- Glycosylation and Glycoproteins Research
- Microbial Inactivation Methods
- Immune Cell Function and Interaction
- Magnetic and Electromagnetic Effects
- Bacteriophages and microbial interactions
- RNA and protein synthesis mechanisms
- Immunotherapy and Immune Responses
- Transgenic Plants and Applications
- Plant tissue culture and regeneration
- Click Chemistry and Applications
- Biochemical and Structural Characterization
- CRISPR and Genetic Engineering
- T-cell and B-cell Immunology
- Viral Infectious Diseases and Gene Expression in Insects
- Advanced Biosensing Techniques and Applications
- Advanced biosensing and bioanalysis techniques
- Platelet Disorders and Treatments
- Receptor Mechanisms and Signaling
- Cancer Immunotherapy and Biomarkers
- Chronic Lymphocytic Leukemia Research
- Protein purification and stability
- RNA Research and Splicing
- Cell Adhesion Molecules Research
Menlo School
2020-2023
Affymax (United States)
1992-2005
Scripps Research Institute
1998
Research Triangle Park Foundation
1997
Baylor University
1995
Stanford University
1979-1994
Stanford Medicine
1994
National Institutes of Health
1980
National Cancer Institute
1980
University of Hawaii System
1975
E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA brief but intense electrical fields exponential decay waveform (electroporation). We have obtained 10(9) 10(10) transformants/micrograms with strains LE392 DH5 alpha, plasmids pUC18 pBR329. The process is highly dependent on two characteristics the pulse: electric field strength pulse length (RC time constant). frequency transformation linear function concentration over at least six orders...
We have constructed a vast library of peptides for finding compounds that bind to antibodies and other receptors. Millions different hexapeptides were expressed at the N terminus adsorption protein (pIII) fd phage. The vector fAFF1, derived from tetracycline resistance-transducing fd-tet, allows cloning oligonucleotides in variety locations 5' region gene III. A 3 x 10(8) recombinants was generated by randomly synthesized oligonucleotides. screened high-avidity binding monoclonal antibody...
The functional mimicry of a protein by an unrelated small molecule has been formidable challenge. Now, however, the biological activity 166-residue hematopoietic growth hormone, erythropoietin (EPO), with its class 1 cytokine receptor mimicked 20-residue cyclic peptide in sequence to natural ligand. crystal structure at 2.8 Å resolution complex this agonist extracellular domain EPO reveals that dimer induces almost perfect twofold dimerization receptor. assembly differs from human hormone...
We have used an in vitro protein synthesis system to construct a very large library of peptides displayed on polysomes. A pool DNA sequences encoding 10(12) random decapeptides was incubated Escherichia coli S30 coupled transcription/translation system. Polysomes were isolated and screened by affinity selection the nascent immobilized monoclonal antibody specific for peptide dynorphin B. The mRNA from enriched polysomes recovered, copied into cDNA, amplified polymerase chain reaction (PCR)...
Two families of small peptides that bind to the human thrombopoietin receptor and compete with binding natural ligand (TPO) were identified from recombinant peptide libraries. The sequences these not found in primary sequence TPO. Screening libraries variants one under affinity-selective conditions yielded a 14-amino acid (Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-Ala-Arg-Ala) high affinity (dissociation constant approximately 2 nanomolar) stimulates proliferation TPO-responsive Ba/F3 cell...
We have prepared a library of approximately 10(6) different peptide sequences on small, spherical (10-microns diameter) beads by the combinatorial chemical coupling both L- and D-amino acid building blocks. To each bead is covalently attached many copies single sequence and, additionally, unique single-stranded oligonucleotide that codes for sequence. The tags are synthesized through parallel procedure effectively records process which encoded assembled. collection was screened binding to...
Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability cell membrane. We investigated utility high-voltage electroporation as a method for genetic transformation intact bacterial using enteric pathogen Campylobacter jejuni model system. This report demonstrates that application discharges to transformation. Our involves exposure suspension exponential decay discharge (5-13 kV/cm) brief period time (resistance-capacitance...
Livers of egg-laying species contain abundant mRNAs encoded by both estrogen-responsive and constitutively expressed genes. We have recently constructed cDNA clones from three members the mRNA class hen liver. One these was identified as serum albumin mRNA, another vitellogenin mRNA. In this study we third member group apo VLDLII Hybridization analyses using cloned probes indicate that expression gene in rooster liver, like gene, is completely dependent upon administration estrogen. The...
E-selectin is an inducible cell adhesion molecule which mediates rolling of neutrophils on the endothelium, early event in development inflammatory response. Inhibition selectin-mediated a possible means for controlling inflammation-induced diseases, and several classes compounds have been tested this use. We describe here use recombinant peptide library screening identification optimization novel ligands bind to E-selectin. Several these peptides with Kd values low nanomolar range block...
Resistance to methotrexate (MTX) has been shown in mouse, hamster, and human cell lines grown sequentially increased MTX concentrations be due dihydrofolate reductase (DHFR) synthesis a proportional increase DHFR gene copy number. Leukemia cells of patient were studied assess change number after treatment. The presented with chronic myeloid leukemia which rapidly evolved into blast crisis; 90% peripheral white lymphoblasts. Treatment included intrathecal intravenous MTX; the lymphoblasts...
The nuclei of the giant neurons marine mollusk Aplysia californica can contain more than 0.2 microgram DNA. This is 200,000 times as much DNA haploid amount found in sperm. On basis nuclear content, R-2, P-1, and L-6 adult animals each be divided into at least two populations. mean content these populations (0.067 0.131 DNA) are approximately related by a factor 2. suggests that perhaps all genome replicates repeatedly (up to 16 times) during growth development replication synchronous....
Two families of peptides that specifically bind the extracellular domain human type I interleukin (IL-1) receptor were identified from recombinant peptide display libraries. Peptides one these blocked binding IL-lalpha to IL-1 with IC50 values 45-140 microM. Affinity-selective screening variants produced ligands much higher affinity (IC50 approximately 2 nM). These block IL-1-driven responses in and monkey cells; they do not II or murine receptor. This is first example (that we know of) a...
To obtain information about the functional importance of amino acids required for effective erythropoietin (EPO) mimetic action, conserved residues a peptide EPO, recently discovered by phage display, were subjected to an alanine replacement strategy. Further, identify minimal sequence, series truncation peptides has been generated. One EPO EMP1, was targeted and more than 25 derivatives this sequence evaluated their ability compete with [125I]EPO receptor binding support proliferation two...
The filamentous phage coat protein pIII has been used to display a variety of peptides and proteins allow easy screening for desirable binding properties. We have examined the biological constraints that restrict expression short located in early mature region pIII, adjacent signal sequence cleavage site. Many functionally defective fusion contained several positively charged amino acids this region. These residues appear inhibit proper insertion into Escherichia coli inner membrane,...
Peptide ligands for the antigen binding site of surface immunoglobulin receptor a human B-cell lymphoma cell line were identified with use filamentous phage libraries displaying random 8- and 12-amino acid peptides. Corresponding synthetic peptides bound specifically to this blocked an anti-idiotype antibody. The ligands, when conjugated form dimers or tetramers, induced death by apoptosis in vitro IC50 between 40 200 nM. This effect was associated specific stimulation intracellular protein...