Chromatography Technique for Purification

Chromatography Technique for Purification Introduction Column packing is an integral part of the purification process in the manufacture of biologics. The goal is always to ensure reproducibility with regard to the technique to be used. Manual packing might sometimes involve several attempts to get an optimal packing as this would affect the purification process. The resin to be used in the packing process has to be well defined as it could impact on the flow rate which could lead to a reduced through put. The mobile phase is also important as the rationale behind the choice would be looking for a solvent that can pack the resin more tightly. In any chromatography technique that has to be utilised whether for the need to capture, purify or polish four integral parameters which includes, resolution, speed, capacity and recovery are always considered. Resolution is the most difficult to achieve especially during the polishing stage were impurities can be construed as having similar properties to the product. The efficiency of the column packing thus has a significant role to play on this basis as it is a good measure how consistently the column can perform. 1.1 Material: As per SOP 1.2 Key Instrument Components Bubble Trap BT1 Filter Housing F1 Inlet / Outlet Valves V001 V101 UV Sensor – QIR4 pH Probe – QIR3 Conductivity Meter – QIR2 Drain – V102 Column top and bottom connections – DN Â ¼ 1.3 Preparation of column for packing: As per SOP TRG-DSP-052. 1.4 Determination of % slurry: Procedure was followed as per instruction manual Results: Table 1 Results: Table 2 r=5cm h= 15cm 1.178 Volume of gravity settled resin for packing (Vgs) 1.178 x 1.33 1.56674 Slurry volume needed from container (SVc) (To give you the desired amount of gravity settled resin) (c) Vgs x 100 % slurry in container = 1.55574 X 100/66 % Slurry in container = 66% 2.37384 Adjusting the slurry to the desired % concentration for packing Slurry volume required for packing (SVp) Vgs x 100 % slurry for packing = 1.56674 X 100/ 70 % Slurry for packing = 70% 2.2382 Volume packing buffer to add SVp – SVc = 2.2382- 2.37384 -0.13564 Volume to be added is thus 0.13564 Litre Calculations: Number of theoretical = N Where VR = volume eluted from the start of sample application to the peak maximum = 8CM W h = peak width measured as the width of the recorded peak at half of the peak height = 0.5CM N = 5.54 X Â ² Number of theoretical plates = 1418.24 HETP = L/N Where L = Bed height (cm) As we already know N (1418.24) HETP = 15/ 1418.24 = 0.0105765 Asymmetry factor (AS) = b/a Where a = 1st half peak width at 10% of peak height (0.5cm) b = 2nd half peak width at 10% of peak height (0.5cm) = 0.5/0. = 1 As rule of the thumb a good HETP value should be at least two to three times the average matrix bead size and normally in the range of 0.0018cm to 0.035cm. Looking at our column our HETP value was approximately 0.0106 and our bead has a pore size of about 40 microns which equates to 0.004cm and this is about 3 times our HETP. Our column can thus be confirmed to be within the acceptable range. In the event that our column is not within the acceptable range several factors such as the following can be construed as being responsible. Uneven packing of the column or exceeding the optimal packing flow rate The possibility of channelling in the bed Inadequate CIP can also be a factor as this can result in a build-up of contamination in the column thus impacting on flow and other performance determinants of the column. Cleaning is also important to wash the matrix storage solution which is an unwanted entity during packing. Air entrapment prevalence of air bubbles can also affect the HETP values. The possibility of a void being present at the inlet can also be a contributing factor to the value of HETP not being within specification The choice of resin is also very important as the possibility of the solute reacting with the resin can result in an ambiguous HETP value. Peak asymmetry is an important measure in the determination of column efficiency and in conjunction with the HETP value is always used in the calibration of a new or existing column. The gold standard is the ability to achieve an asymmetry value of 1 although the acceptable range is normally between 0.8 and 1.2. An asymmetry value greater than 1 indicates the prevalence of extensive tailing while an asymmetry value less than 1 indicates extensive fronting. Taking our packed column into consideration, our asymmetry value from the chromatogram was 1 and one would generally thus expect a high efficiency and resolution. However, in the event of our column not being within the acceptable asymmetry value the following reason are the possible causes. Extensive tailing which is characterised by an asymmetry value greater than 1 as mentioned earlier can be a reason. This factor is a result of column being packed too loosely and it can be observed from the chromatogram by the peak tailing gradually. Extensive fronting is also a possible cause and it is characterised by an asymmetry factor less than 1 which is normally as a result of the column being packed tightly and would be noticeable on the chromatogram by the peaks developing slowly. Possible causes of resin/column deterioration and their remedies Temperature the resins have a temperature range that is normally specified by the manufacturers and a usually high temperature can cause irreversible damage due to loss of functional groups. It is thus important that operation should always within the optimal ranges and bearing in mind the fact that temperature maxima is only for indication. Oxidation – The functional groups are also attached by oxidation and on this basis one has to ensure that oxidants such as hydrochloric acid , nitric acid are not utilised in the cleaning regime as they can accelerate oxidation which damages the polymer crosslink Fouling apart from impacting on performance of the column can also cause irreversible damage to the resin. Fouling can result due to the presence of iron and silica for this reason special attention has to be paid to the type of resin to be used as prevention they say is better than cure. Drying out and cracking of the resin is also an important reason for column deterioration and this can be remedied by ensuring that the column is well equilibrated. High pressure -The build-up can also cause damage to the resin/column and it could be as a result of flow path restriction due to dirty or worn bed support. Manufacturer’s specification should always be adhered to in ensuring an optimal usage of the resin. The life span of the resin/column should also be taken into account and usage should always be as specified by the manufacturer. Harsh elution is another factor that is responsible for irreversible damages to resin/column. Every resin has a pH range that is optimal and this should be adhered to strictly. Conclusion: The practical experience was so interesting and brought the protein purification lectures received into perspective. A better understanding of the process was developed and the practical knowledge is quite adaptive to the day to day operation in a typical Biopharmaceutical plant.

Symbols and Symbolism - Flowers as a Symbol in John Steinbecks The Chr

Flowers as a Symbol in The Chrysanthemums by John Steinbeck      Ã‚  Ã‚   In romantic or sexual context, a woman is often said to be as beautiful as a flower. In John Steinbeck's short story "The Chrysanthemums", Elisa Allen never receives this recognition. Although she is a strong woman, she is frustrated because her husband will not admire her romantically in any way. This frustration only deepens because she is childless and feels the need to be a mother. She discovers an outlet for her frustration in a flower garden where she cultivates beautiful chrysanthemums. Steinbeck uses these elegant flowers as a symbol to represent the tender, inner-self of all women, including Elisa. First, the chrysanthemums symbolize Elisa's children. She tends her garden and handles the chrysanthemums with love and care, just as she would handle her own children. Elisa is very protective of her flowers and places a wire fence around them; she makes sure "[n]o aphids, no sowbugs or snails or cutworms" are there. "Her terrier fingers [destroy] such pests before they [can] get started" (240). These pests rep... ...s and her emotional needs. The encounter with the tinker reawakens her sexuality and brings hope to Elisa for a more exciting and romantic marriage, but her realization that her life is not going to change is crystallized when she sees the flowers thrown on the road. It devastates her completely to have to settle for such an unfulfilling life. Work Cited Steinbeck, John. "The Chrysanthemums." Literature: An Introduction to Fiction, Poetry, and Drama. Ed. X. J. Kennedy and Dana Gioia. 6th ed. New York: Harper Collins, 1995. 239-47.  

Personality Development Analysis Essay

Personality development is among the important aspects of life that we have to go through to ensure a complete and healthy life mentally, physically and emotionally. In this paper the three major theories of personality development are going to be used to analyse an individual that has already been chosen. Erickson’s theory of psychosocial development, Piaget’s theory of cognitive development, and Freud’s theory of psychosexual development are going to explain the various traits of our character in an effort to try and further understand the human personality development. The character has been chosen from the television series ‘Scrubs’; he is Dr John Michael Dorian also known as ‘JD’. JD is a 32 year old medical doctor; currently he is the residency director at St Vincent’s hospital, a position he recently acquired. He can be classified as a genuinely good doctor; this is shown by his concern for his patients while he was working at Sacred Heart Hospital. His best friend is Dr Chris Turk who is the Chief Surgeon at Sacred Heart hospital. The two of them have been together for most of their adult life; it has been frequently been confused that they are homosexual because of the closeness of their relationship. JD has a son called Sam who he adores. He has a girl friend called Elliot also a doctor in private practice. His mentor is Dr. Perry Cox who is the current chief of medicine at Sacred Heart hospital. All of them have worked together for over 8 years; they all have various close relationships depending on their line of work, perceptions e. t. c. Lastly JD has been portrayed as a needy human being; this is shown by his sensitivity and neediness. He has been described as the neediest person in the hospital. According to Erickson’s Psychosocial Development theory, JD is in the Intimacy vs. Isolation stage. At this stage the main virtue is love and understanding. It is at this stage that one dares to experience relationships with other people. These relationships are therefore key aspects in ones life at this stage in life (Feldman 2005). When we look at JD, we see that he has a network of relationships that actually determine how well he works as a doctor. His relationship with Dr. Turk is of more importance. The call each other using pet names; brown bear and vanilla bear, this shows that they are both comfortable with each other. This relationship is also characterised with a lot of child like games and competitions. The two can therefore be considered as inseparable no matter what the circumstances (Dacey & Travers 2002). They depend on each other through thick and thin. There is also a lot of focus on JD career. He has been described as the residency director at St Vincent’s Hospital a position that he accepted because he would like to live near his son; he wants to spend more time with his on Sam. That is another relationship that dominates his life, in fact since both JD and Turk have children, when ever they get free time they spend it together with their children. This shows that the relationship between the parents and the children is healthy and will be healthy in the future, another characteristic that JD is in the Intimacy vs. Isolation stage. Unfortunately, JD has had a series of bad relationships with the opposite sex; this attributed to the fact that he is not emotionally healthy at that level, very small things with the opposite sex tends to bring out the worst in him. This is not for lack of trying but because as he defines it he self sabotages all his romantic relationships, a trend that can be seen also in his social life though with varying degrees. That is why he is mostly lonely (Dacey & Travers 2002). According to the Piaget’s theory of cognitive development, JD as a rational adult is in the Formal Operational stage. He is able to think rationally and therefore reach rational conclusions. He can speak eloquently and therefore an effective communicator. He knows the difference between right and wrong; it is part of the reason as to why he is a good doctors. According to JD patients need the extra attention so that they can heal even faster. This he reached despite constant ridiculing by his friends and at time even the doctor. But he still feels that it is right to spend sometime to know ones patient. Some of the skills that one acquires when at this stage have also been demonstrated; JD uses his deductive skills to diagnose his patients (Leifer & Hartston 2004). This also enables him to spend more quality time with his patients. It is the same reason as to why he is very much attached to the same patients. He is able to make rational decisions; seen when he decides to take another job so that he can spend more time with his son. This is particularly a hard decision because it means moving away from his best friend Turk. It means spending less and less time with Turk; many still don’t believe JD could make such a decision, but he does. This shows that he understands that in life one has to learn how to make rational decisions that will further enhance our lives and the lives of our loved ones. Being selfish is therefore a sign that all is not well. Life sacrifices have to be made so as to ensure that we are with those that we love and cherish. Such sacrifices come out of making rational decisions and although some may be sad, one has to look at the bigger picture. On the other hand JD is know for making the wrong decision despite knowing what the consequences are; he has an inner monologue that is constantly telling him what he should do but he always ignores this advice only to suffer the consequences latter (Leifer & Hartston 2004). According to Freud’s theory of Psychosexual Development, JD has successfully gone through the required stages that one has to go through to ensure that he is ready and complete for life. He is an adult and has therefore managed to go through all the processes as described by Freud; he is therefore at the genital stage. At this point in life JD is struggling to establish a balance between a lot of things going on in his life. He therefore reaches a decision that favours him moving a way from what he is used to so as to be with his son. He also makes a decision that involves him living with Elliot his girlfriend and eventually his wife. At this stage one tends to focus on the needs of others rather than their own needs and this explains why JD made such a selfless decision so that he can be with his wife and further enhance his career as a residency director. At his point in life one also tends to be attracted to the members of the opposite sex; the decisions at this point could be self serving or simply just wrong but a relationship borne out of such a decision in this stage rarely lasts as one discovered that they were only attracted to the person because of one thing, their sexuality (Feldman 2005). In an analysis of all the theories, then it is safe to conclude that JD is in fact a normal healthy human being compared to any standards. Despite a few flaws here and there, he is emotional stable and mentally sound. He is a hard working member of the community and in no way a perfect person. A balance is therefore necessary; a balance of all the elements of personality development. This balance means that one is tolerable and consequently of sound abilities. JD has friends and family around him who love him and appreciate him; his childish nature has been described as a very normally way to avoid getting overwhelmed since he is in a demanding career. The fact that he is father means that he is a man who will do everything to ensure that his son never lacks anything and hopefully. In conclusion the theories of personality are meant to try and establish criteria through which a human being develops; they try and explain why human personalities are different and at times we all happen to go through the same things in life. Psychosocial characteristics are meant to establish how and why a person relates to others the way they do. It explains person’s social characteristic within the social context. Cognitive development explains tries to understand how humans acquire knowledge and therefore deduces a person’s personality through this process of learning. In the end we have the psychosocial development that is mainly focused on the sexual drive in human beings and how people’s personalities develop as a result of undergoing these urges; a balance and control of these urges at different stages in life means that one will end up with a good personality. But in the end a balance of almost all elements remains desirable; depending on the theories a balance of all these attributes means that one is of good nature and therefore a good human being to be with. References: Dacey John S & Travers John F. (2002). Human Development across the Lifespan. Boston: McGraw-Hill. Feldman Robert S. (2005). Development across the life Span, Upper Saddle River, N. J: Prentice Hall. Leifer G & Hartston H. (2004). Growth and Development across the Lifespan: A health Promotion Focus. St. Louis, Mo: Saunders.

The Crucible - Comparing Play and Movie - 1004 Words

#9;Arthur Miller s play, The Crucible, and the movie with the same name have many differences and similarities, all of which contribute to the individual effectiveness of each in conveying their central message. #9;There are several additions and variances in the movie. First of all, the scene where the children and Tituba are dancing in the forest is never seen in the forest; we simply learn of it from dialogue. This scene was most probably added in the movie for a dramatic effect: foreshadowing. Along the same lines, there are many scenes included in the movie that are outdoors, however; there were no such scenes in the play. All of the action in the play was indoors. The director, in this case, was simply using the advantages of†¦show more content†¦I believe the purpose of their running into the ocean was that the girls needed to cleanse themselves of the evil spirits. The accusation of John Proctor practicing witchcraft comes when he is standing in the water to his mid-thigh, possibly meaning that, although he has sinned less than some, he still needs to be baptized (in a sense) completely to be free of his misdoings. The final three differences all have to do with Abigail. First of all, she is seen stealing money from her uncle where in the play we only hear of it second-hand. Second, in the movie, Abigail visits John Proctor in jail, which never occurred in the play. Lastly, Abigail accuses the Reverend John Hale s wife of witchcraft, and is told by Judge Danforth that she is mistaken. This did not happen in the play, but helps in the movie to more clearly define Abigail as one of the bad guys in the movie. Personally speaking, I prefer the movie to the play. Granted, the play may have deep, underlying meanings, but to me, it wasn t very moving or emotional. The additions made in the movie help to more clearly define the roles of good and evil, and play on the hidden feelings people have. I think most of the additions, if not all, were appropriately made and were quite successful. I enjoyed both the movie and our reading of the play very much, but again, I would have to say that I prefer theShow MoreRelated The Crucible - Comparing Play And Movie Essay974 Words   |  4 Pages Arthur Millers play, The Crucible, and the movie with the same name have many differences and similarities, all of which contribute to the individual effectiveness of each in conveying their central message. There are several additions and variances in the movie. First of all, the scene where the children and Tituba are dancing in the forest is never seen in the forest; we simply learn of it from dialogue. 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